Bi-functional co-polymer use for ophthalmic and other topical and local applications

ABSTRACT

The invention contemplates a copolymer which is a graft or block copolymer useful to change the ocular surface temperature and other characteristics of biological or contact lens surfaces. Methods for use of these formulations and coatings to increase the temperature of the skin, mucous membranes, eye or eyelids will help treat many conditions including blepharitis and non-healing ulcers. Methods to decrease evaporation, improve wettability and stabilize the tear film, and lubricate biological surfaces in a subject, for example, in the treatment of dry eye syndrome, and to improve contact lens tolerability, are provided.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.16/241,147 filed Jan. 7, 2019, which is a continuation of internationalapplication No. PCT/US2016/041079, filed on Jul. 6, 2016, the entirecontents of each of which are incorporated herein by reference.

FIELD OF THE INVENTION

The present invention provides topical and/or local deliverypharmaceutical formulations and coatings for extraocular devices, drugparticles, or drug containing particles comprising a bi-functionalpolymer or co-polymer and methods for their use in modifying surfacecharacteristics and/or changing the wettability of biological surfacesin a subject, or on the aforementioned devices. Formulations of thepresent invention are particularly useful in various applicationsincluding increasing the ocular surface temperature, preventingevaporation, hydration, in particular treatment of dry eye syndrome andtreatment of ophthalmic discomfort prevention of infection and/oradhesion of bacteria, viruses, proteins, toxins, and antigens anddelivery of active pharmaceutical ingredients to biological surfaces.Formulations of the present invention can be used to alter thetribological properties of biological surfaces. Formulations of thepresent invention are also useful when combined with a host of topicallydelivered therapeutic agents. The formulations are also useful forcoating extraocular devices, particularly contact lenses, to improvetheir biological compatibility.

BACKGROUND OF THE INVENTION

Poly(L-lysine)-graft-poly(ethylene glycol) (PLL-g-PEG) is a watersoluble co-polymer consisting of a poly(L-lysine) backbone andpoly(ethylene glycol) side chains (Sawhney et al. Biomaterials 199213:863-870). The PLL chain, which carries multiple positive charges,spontaneously adsorbs onto negatively charged surfaces while PEG is apolynonion which serves as a non-binding domain. This adsorption isstrong and occurs rapidly, and renders surfaces protein and cellresistant. Furthermore, PLL-g-PEG has been shown to improve thebiocompatibility of materials. (Sawhney et al. Biomaterials 199213:863-870)

Various applications for the PLL-g-PEG graft co-polymer, blockco-polymers, and dendrimers with similar binding characteristics havebeen described.

SUMMARY OF THE INVENTION

In one aspect, the invention provides a method for increasing thewettability of a biological surface or treating biological surfacerelated discomfort in a subject, involving contacting the surface with acomposition containing a copolymer having a positively charged, orhydrophobic or covalent bonding moiety and a hydrophilic moiety, wherethe contacting is effected in an amount and for a duration so as toincrease the wettability or treat discomfort of the biological surface.

In one aspect, the invention provides a composition containing acopolymer having a positively charged, or hydrophobic, or covalentbonding moiety and a hydrophilic moiety and a pharmaceuticallyacceptable carrier.

In one aspect, the invention provides a pharmaceutical formulationcontaining an effective amount of a composition containing a copolymerhaving a positively charged, or hydrophobic, or covalent bonding moietyand a hydrophilic moiety and a pharmaceutically acceptable carrier.

In another aspect, the invention provides a method of storing a contactlens for at least 1 hour, involving providing the contact lens and apharmaceutical formulation of the invention.

In one aspect, the invention provides a kit containing a contact lensand a pharmaceutical formulation of the invention, in a container thatis unopened subsequent to manufacture.

In one aspect, the invention provides a kit containing a sterilepharmaceutical formulation of the invention and packaging materialstherefore which permit use as an artificial tear product introducibleinto the eye.

In one aspect, the invention provides a kit comprising an extraoculardevice coated with the composition of the invention, and packagingmaterials therefore. In various embodiments, the packaging materialspermit contacting of the extraocular device through the nozzle of an eyecare solution container. In various embodiments, the extraocular deviceis an intracorneal inlay.

In one aspect, the invention provides a method for manufacture of acontact lens, involving providing a contact lens and contacting thecontact lens with a composition of the invention.

In another aspect, the invention provides a kit containing a contactlens and a pharmaceutical formulation of the invention, in a containerthat is reusable.

In one aspect, the invention provides a copolymer coated nanoparticle,the surface of the nanoparticle containing a copolymer having apositively charged, or hydrophobic, or covalent bonding moiety and ahydrophilic moiety. In various embodiments, the nanoparticle furthercontains an active agent.

An aspect of the present invention relates to a method for changingwettability of and/or sterically modifying a biological surface in asubject comprising locally administering to the biological surface ofthe subject a pharmaceutical formulation comprising a bi-functionalco-polymer. A copolymer composition can be contacted with an externalbody surface (e.g., skin) and/or used to prevent infection. Ofparticular interest and focus in this invention is the external surfaceof the eye including bulbar and palpebral conjunctiva and the cornea.Another aspect of the present invention relates to a method for treatingdry eye syndrome comprising topically administering to an eye of asubject with dry eye syndrome a pharmaceutical formulation comprising abi-functional co-polymer.

Another aspect of the present invention relates to a method forprotecting a biological surface in a subject against pathogen or toxinattack, said method comprising topically or locally administering to thebiological surface in the subject a pharmaceutical formulationcomprising a bi-functional co-polymer.

Another aspect of the present invention relates to a method fordelivering an active pharmaceutical ingredient to a biological surfacein a subject comprising topically or locally administering to thebiological surface of the subject a pharmaceutical formulationcomprising a bi-functional co-polymer and the active pharmaceuticalingredient.

Another aspect of the present invention relates to a formulation for usein methods of changing wettability of (or otherwise sterically modifyingor stabilizing)a biological surface in a subject, treating or preventingdry eye syndrome, treating or preventing discomfort relatedcomplications of LASIK surgery, treating or preventing infectious orinflammatory conditions of the external eye, protecting a biologicalsurface in a subject against pathogens (bacteria, fungi, virus, prions,bioadhesive chemical agents) and/or delivering an active pharmaceuticalingredient to a biological surface in a subject, said formulationcomprising a bi-functional co-polymer for topical and/or localadministration to the subject.

Another aspect of the present invention relates to coating extraoculardevices with a bi-functional co-polymer. Another aspect of the presentinvention relates to the use of the bi-functional co-polymers describedherein to prevent or treat contact lens related discomfort and to reducesymptoms of contact lens intolerance.

Other aspects of the invention include the coating and thus protectionof biological surfaces, such as the skin, especially mucous membranesinside the mouth, nose, throat, ear, vagina, for the purpose ofmaintaining hydration, and for protecting the biological surface frominfection by pathogens and from toxins.

Other aspects of the invention include coating drug particles orparticles containing drug to increase their utility by increasinghalf-life and/or decreasing interaction with biologic phenomenon such asan immune response.

In various embodiments of any of the above aspects, the copolymer is agraft or a block copolymer. In various embodiments, the copolymer is agraft copolymer and contains a cationic backbone and side chains thatare water soluble and non-ionic. In various embodiments, the copolymeris a graft copolymer and contains a water soluble non-ionic backbone andcationic side chains. In various embodiments, the copolymer is a blockcopolymer and contains at least one cationic block and at least onewater soluble and non-ionic block. In various embodiments, the copolymeris a block copolymer and contains at least one block which ishydrophobic and at least one block which is water soluble and anionic.In various embodiments, the copolymer is a block copolymer and containsat least one block which is hydrophobic and at least one block which iswater soluble and cationic. In various embodiments, the graft copolymeris PLL-g-PEG.

In various embodiments of any of the above aspects, the copolymer is adendrimer. In various embodiments of any of the above aspects, thehydrophilic moiety is one of non-ionic or anionic. In variousembodiments of any of the above aspects, the copolymer is a blockcopolymer and contains at least one block which is hydrophobic and atleast one block which is water soluble and or non-ionic.

In various embodiments of any of the above aspects, the copolymercreates a covalent adhesion between the copolymer and the biologicalsurface. In various embodiments of any of the above aspects, thecontacted biological surface is the surface of an eye, a mucousmembrane, or skin of a subject.

In various embodiments of any of the above aspects, the copolymer is0.001 to 40% of the composition. In various embodiments of any of theabove aspects, the copolymer is 0.001 to 25% of the composition.

In various embodiments of any of the above aspects, the compositionfurther contains a second polymer. In various embodiments of any of theabove aspects, the composition further contains a PLURONIC blockcopolymer. In various embodiments of any of the above aspects, thecomposition further contains one or more of a surfactant, apreservative, and a pharmaceutical ingredient including demulcents[Cellulose derivatives (e.g., Carboxymethylcellulose sodium,Hydroxyethyl cellulose, Hydroxypropyl methylcellulose, Methylcellulose),Dextran 70, Gelatin, Polyols (e.g., Glycerin, Polyethylene glycol 300,Polyethylene glycol 400, Polysorbate 80, Propylene glycol), Polyvinylalcohol, Povidone] emollients (e.g., Lanolin, mineral oil, paraffin,petrolatum, white ointment, white petrolatum, white wax, yellow wax),sodium chloride, and vasoconstrictors (e.g., Ephedrine hydrochloride,Naphazoline hydrochloride, Phenylephrine hydrochloride, andTetrahydrozoline hydrochloride).

In various embodiments of any of the above aspects, the composition iscontacted to the biological surface in an amount sufficient to changetribological properties of the biological surface of the subject. Invarious embodiments of any of the above aspects, the composition iscontacted topically to an eye of a subject. In various embodiments ofany of the above aspects, the composition is topically administered toan eye of a subject to treat dry eye, ophthalmic irritation, or cornealepithelial disease. In various embodiments, the composition is contactedto the eye by an eye drop or an eye care solution. In variousembodiments, the method further involves administering to the eye of thesubject a second, different eye drop. In various embodiments of any ofthe above aspects, the composition has a low viscosity artificial tearless than 20 cP, having prolonged tear film break up time. In variousembodiments of any of the above aspects, the positively charged moietyis sufficient as a preservative in a topical ophthalmic formulation. Invarious embodiments of any of the above aspects, composition is in avolume sufficient for instillation in the eye, wherein the copolymer isPLL-g-PEG at a concentration of 0.1 to 3 wt %.

In various embodiments of any of the above aspects, the copolymer isimmobilized on an extraocular device at the time of manufacture. Invarious embodiments, the copolymer is immobilized on the extraoculardevice covalently. In various embodiments, the immobilization iseffected via an aldehyde plasma polymer interlayer and reductiveamination. In various embodiments, the extraocular device is a contactlens.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 shows results from an in vitro assessment of the ability ofexemplary formulations of the present invention containing PLL-g-PEG todecrease contact angle of water to polystyrene. Formulations containing10 mg/ml or 1% (closed circles), 5 mg/ml or 0.5% (open circles) or 1mg/ml or 0.1% (closed triangles) were compared to a control of deionizedwater (open triangles).

FIG. 2 shows results from an in vitro assessment of the ability ofexemplary formulations of the present invention containing F87 or F127to decrease contact angle of water to polystyrene. Formulationscontained either 1% F127 (open circles) or 1% F87 (closed circles) andwere compared to the average contact angle of deionized water (straightline).

FIG. 3 shows the results of a randomized, controlled, double maskedclinical trial in sixteen subjects where the tear film break-up time wasmeasured at varying time points after eye drop instillation. Thisclinical methodology is a common assessment tool for dry eye syndrome,as well as to determine the effectiveness of artificial tears. Fifteenminutes after instillation of sample formulation and active control,sample formulation showed extension of noninvasive tear film break-uptime from baseline of 14.67 sec (p=0.05), while active comparator was7.4 seconds longer than baseline (p=0.34).

FIG. 4 shows results from the same randomized, controlled, double maskedclinical trial where the tear film break-up time was measured at varyingtime points in subjects. Fluorescein break-up time was longer thanactive comparator at 120 minutes; superior by 4.92 sec (p=0.12).

FIG. 5 shows results of a sub group analysis of the trial describedabove. In the main group of 16 subjects in the trial, there were threesubjects that had unexpectedly long pre-instillation TFBUT. These longinitial values distorted the relative contributions of the artificialtears to NIBUT for the other subjects. When these three outliers wereremoved, there was superiority in extending NIBUT for the sampleformulation at every time point. The data is graphed based on extensionby seconds over baseline in FIG. 5.

FIG. 6 shows results of a sub group analysis of the trial describedabove similar in method to FIG. 5. In the main group of 16 subjects inthe trial, there were three subjects that had unexpectedly longpre-instillation TFBUT. These long initial values distorted the relativecontributions of the artificial tears to NIBUT for the other subjects.When these three outliers were removed, there was superiority inextending NIBUT for the sample formulation at every time point. The datais graphed by percentage change from baseline in FIG. 6.

FIG. 7 is an example schematic drawing of the molecular behavior of thegraft copolymer on an epithelial or ophthalmic device surface. Thisschematic demonstrates how the surface modifying polymer can bebeneficial in altering the wettability or tribological properties of asurface and/or sterically modifying the surface so that the epitheliumis protected and stays wet longer, or how an extra- or intra-oculardevice may be made more biocompatible and less likely to foul withproteins and cellular debris. 701 represents the negatively chargedepithelial surface (such as the cornea and conjunctiva) or negativecharges on a device (such as a contact lens). 702 represents thepositive charge on the NH3 terminal of lysine. 703 represents theanchoring poly (L) lysine backbone. 704 represents the flexiblepolyethylene glycol (PEG) side chains that form a brush. This brush canimpart protein resistance, and is hydrated (705 is a water molecule),and thus there is also a change in wettability imparted onto the surfaceon which the graft (or block) copolymer electrostatically (or throughhydrophobic interaction in some cases) adheres. 706 represents theentire graft copolymer molecule (PLL-g-PEG in this example). Stericstabilization is also imparted to exposed surfaces decreasing contactwith proteins and cells, thereby protecting the epithelial or devicesurface from protein or cellular adhesion, inflammation or fouling. Thisfigure is an example of PLL-g-PEG, but the same concept applies forother graft copolymers, generally, using either hydrophobic or cationicinteraction with a surface combined with a hydrophilic moiety impartingthe protective aspects of the invention herein described. Furthermore,the bi-functional co-polymers, block copolymers, and dendrimers may bediagrammed in a similar fashion, using a repeating moiety adherent tothe surface and a repeating hydrophilic moiety imparting changes inwettability and protection. Furthermore, in some embodiments, thebi-functional copolymer may be covalently immobilized on the surface ofa device such as a contact lens.

DETAILED DESCRIPTION OF THE INVENTION

It has now been found that topical or local administration of aformulation comprising a graft co-polymer having a positively charged orhydrophobic moiety and a hydrophilic moiety or a block co-polymer havinga positively charged or hydrophobic moiety and a hydrophilic moiety to abiological surface alters and increases the wettability of certainsurfaces, including a biological surface particularly the corneaconjunctiva and external surface of the eye, and does so for a prolongedperiod of time. A copolymer composition of the invention can becontacted with an external body surface (e.g., skin) and/or used toprevent infection.

By “moiety” it is meant the required portion of the polymer. Forexample, the moiety may be a monomer or one aspect of the polymer thatimparts some characteristic required for activity and/or result. The PEGmoiety imparts hydrophilicity. The PLL moiety imparts cationic charges.A phenylboronic moiety for example imparts an ability to form covalentattractions.

By “covalent” it is meant an interatomic bond charcterised by thesharing of electrons. The bond can be reversible or permanent.

By “electrostatic” it is meant interaction between two molecules or twopolymers or between a molecule or polymer and a biological ornon-biological surface based on charge attractions—negative charge onone aspect of entity and positive charge on one aspect of the other.

By “bioadhesive” is meant a synthetic or natural polymer which binds tobiological substrates such as mucosal membranes. Binding may beaccomplished by electrostatic interaction, covalent attraction,hydrophobic interaction or other means.

By “hydrophobic interaction” it is meant an interaction betweenlipophilic moieties on polymers and surfaces to form adhesions orintermolecular aggregates or intramolecular interactions (particularlyin aqueous based environments).

By “adhesive”, it is meant tending to adhere, stick, or not come easilyloose, or tending to persist.

By “increases the wettability” it is meant an increase in the relativedegree with which an aqueous liquid will spread onto or coat a surface,including in the presence of other immiscible liquids. Increasedwettability, according to the invention results in prolonged stabilityof the tear film keeping the cornea better coated by natural andartificial tear film components, reduce adhesion of inflammatoryproteins, bacteria and inflammatory proteins to a biological surface ormembrane of the subject.

Changes in wettability, however, are only part of the advantagesconveyed by this invention. Surface modifications also include thesteric stabilization of biological surfaces and extra-ocular ophthalmicdevice surfaces.

By “biological surface” is meant the surface of a bodily organ, whetherit be exposed to the external environment, or internal to the body. Forexample, the surface of the eye includes the epithelial covered corneaand bulbar and palpebral conjunctiva, as well as the posterior tenonslayer and sclera; the epithelial layers of the gastrointestinal tract orthe skin are included, membranes such as mucous membranes, includingoral, nasal, respiratory, urinary tract including the bladder, andvaginal mucous membranes. Other surfaces include the capsules of organssuch as the spleen and liver, and the outermost aspect of bone,cartilage, and muscle, as well as wounds and areas where there isunprotected subepithelial biological tissue exposure.

By “tribological properties” it is meant properties of interaction ofsurfaces in relative motion. For example, a contact lens on the surfaceof the eye with some movement on the corneal surface and/or with the actof blinking whereby the palpebral conjunctival surface moves over thecontact lens constitutes a state of relative motion between twosurfaces. Likewise, blinking whereby the palpebral conjunctival surfacemoves over the cornea and bulbar conjunctiva constitutes a state ofrelative motion between two surfaces. Tribological properties caninclude but are not limited to properties such as lubrication, friction,and wear.

By “subject”, as used herein it is meant to be inclusive of all animalsand in particular mammals such as, but not limited to, humans and dogsas well as agricultural animals such as bovine, ovine, and porcine.

By “prolonged period of time” means that the formulation has an effectthat lasts longer than leading artificial tear products on the market,which has been shown to be at least two hours with the sampleformulation, but “prolonged period of time” can also mean longer than 20minutes, as most ophthalmic topical products are washed out of the eyein 20 minutes. A “prolonged” period of time includes 30, 45, 50 minutes,as well as 1, 2, 5, 10, 24 hours, or 1, 2, 5, 10 or 30 days or more.

By “steric stabilization” or “steric modification” means that thesurface effects of the graft and block co-polymers results in ahydrophilic moiety projected away from the biological surface, drugparticle or drug containing particle that imparts characteristics suchas low protein adsorption and cell adhesion. Such an effect can also becalled steric repulsion and surface exclusion effects secondary to thePEG moieties (termed “brushes” in some descriptions) on the graftcopolymers. Other hydrophilic chains can accomplish the same purpose insome embodiments. High surface water retaining capacities, osmoticrepulsion, and charge neutrality may also contribute to this “stericstabilization” effect.

By “storage” is meant time on the shelf in a container, and storage timeincludes 1, 2, 5, 10, 30, 180, or 360 days or more.

Accordingly, the present invention provides methods for changingwettability of (or sterically stabilizing or otherwise modifying)biological surfaces or membranes in a subject via topical or localadministration of a pharmaceutical formulation comprising a graftco-polymer having a positively charged or hydrophobic moiety and ahydrophilic moiety or a block co-polymer having a positively charged orhydrophobic moiety and a hydrophilic moiety to a biological surface of asubject, or both. Pharmaceutical formulations can be administeredtopically or locally in accordance with methods of the present inventionto biological surfaces of a subject including, but not limited to, skin,mucous membranes, hair, and the surface of the eye.

According to the present invention, through the use of the block andgraft co-polymers described herein, certain drugs and activebiopharmaceutical agents will have improved efficacy and decreasedfrequency of dosing needs as formulations using the block or graftcopolymers can bring such active pharmaceutical agents into contact withepithelial surfaces, such as the cornea and conjunctiva, for prolongedperiods of time.

According to the present invention, through the use of the block andgraft co-polymers described herein, the tolerability of extraoculardevices such as contact lenses, shunts, retinal implant/explantmaterials and devices for scleral buckles, and drug delivery devices,can be improved. Furthermore, ophthalmic sutures may be coated withthese polymers decreasing tissue reaction and reducing unwanted particleor bacterial adherence. First, the devices can be coated (usingelectrostatic or hydrophobic interactions or through covalentimmobilization), with the said polymers increasing wettability,modifying surface characteristics, and/or sterically stabilizing anddecreasing the adherence of unwanted antigens, cells, proteins, andother particles to them. Second, by protecting the epithelial surface orother ophthalmic tissues (through turnover and/or exchange of theseadherent polymers with a biological surface and/or through theseparation of the device from the epithelial surface by the “brush”characteristics of the hydrophilic moieties), the devices may be moretolerable.

By “extraocular device” is meant medical devices that do touch but donot reside entirely inside the eye. A contact lens is an extraoculardevice. An intra-corneal inlay is an extraocular device because it doesnot breach the intraocular environment. A glaucoma shunt, is anextraocular device because the vast majority of the device (except forthe distal tip of the tube) resides outside the eye. A scleral buckle isan extraocular device.

By “co-polymers” is meant a polymer with two or more different monomers.Block, graft, and dendrimer co-polymers are the major, but not onlytypes of co-polymers referenced herein.

By “bi-functional” is meant the co-polymer has two or more differentfunctions that derive from the different moieties or monomers inherentto the co-polymer. For example a hydrophilic moiety or monomer has theproperty of wettability and a cationic backbone has the property ofbioadhesiviness. Joined together in a co-polymer both functions remain.Specifically, bifunctional co-polymers relates to but is not limited tograft, block, a and dendrimer co-polymers. Critical to this invention isthe novel use of bi-functional graft copolymers in ophthalmic use.

By “ophthalmic use” is meant used on or in or adjacent to the eye.Topical eye drops are included in ophthalmic use as is subconjunctival,peribulbar and eyelid use. Use on contact lenses and extraocular devicesis included in ophthalmic use.

By “pluronic” is meant poloxamers or trade name pluronic copolymers thatare composed of nonionic triblock copolymers composed of a centralhydrophobic chain of polyoxypropylene (poly(propylene oxide)) flanked bytwo hydrophilic chains of polyoxyethylene (poly(ethylene oxide))

By “preserved” it is meant there is a substance or preparation added toa product to destroy or inhibit the multiplication of microorganisms.

According to the present invention, through the use of the block andgraft co-polymers described herein, the tolerability of extraoculardevices (such as contact lenses and intracorneal optical devices), canbe improved. First, the devices can be coated with the said polymersincreasing wettability and decreasing the adherence of unwantedantigens, cells, proteins, and other particles to them. Second, byprotecting the adjacent tissue structures, the devices may be moretolerable and may function better i.e. through less protein or cellularinteraction on the device. In the case of intracorneal inlay technology,such a coating may improve the clinical performance of such devices inmultiple ways.

Graft co-polymers used in the methods and formulations of the presentinvention are polymers having a linear section of repeat units calledthe “backbone”, with at least one side chain of repeat units (called agraft), usually of a different chemistry, branching from a point alongthe backbone. In one embodiment, the graft co-polymer comprises acationic backbone and side chains that are water soluble and non-ionic.In another embodiment, the graft copolymer comprises a water solublenon-ionic backbone and cationic side chains.

Block co-polymers used in the methods and formulations of the presentinvention are polymers in which linear sections of a first section ofrepeat units are connected end-to-end with linear sections of subsequentrepeat units that are chemically dissimilar to the first.

Dendrimers comprised of cationic and hydrophilic polymeric moieties cansimilarly be used as identified above in the methods and formulations ofthe present invention to accomplish similar structural and stericalterations of the biological surface (particularly the eye), drugparticles or drug containing particles.

By “dendrimers” it is meant co-polymer molecules that are repeatedlybranched.

Also included in the invention is the use of graft, block, anddendrimeric copolymers with multi-functionality in topical applicationsand formulations including ophthalmic applications and formulations. Forexample, a polymer where there is a moiety bonded to one constituentpolymer that promotes covalent bond formation between the copolymer andbiological surfaces such as the surface of the eye or between thecopolymer and extraocular devices that also project a comb or a brushtype of hydrophilic moiety, is claimed. As can be seen to one skilled inthe art, the results accomplished with the use of electrostaticinteraction between the polymer and biological surfaces and the externalenvironment or extraocular devices can be accomplished using similarlydesigned bi- and multi-functional polymers that promote the formation ofa covalent bond instead of or in addition to electrostatic attractiononly. For example, the use of phenylboronic acid (PBA) or other boronbased moieties in the polymer backbone provide a method for theformation of covalent complexes between copolymers and biologicalsurfaces and between copolymers and extraocular devices. Even though notcalled out continuously throughout this document, the method of usingcovalent, electrostatic and hydrophobic attraction (or any combinationor either) in ophthalmic applications is described. PLL-g=(PEG; PBA) isan example of a polymeric structure that would impart the describedcharacteristics.

Formulations for use in the methods of the present invention comprise ablock or graft co-polymer having one section, either the backbone, thegraft or the block, that adheres to a biological surface tissue such as,but not limited to, the eye surface by electrostatic or hydrophobicforces (or covalent), and another, chemically different section, eitherthe backbone, the graft, or block, that is hydrophilic and retainsmoisture (sometimes called a “brush” or a “PEG brush” in someembodiments), or allows an aqueous film to readily spread over and thuswet the biological surface. Sometimes the graft copolymers such asPLL-g-PEG, for example, can be referred to as comb co-polymers.Furthermore, the “brush” that is composed of water soluble polymers thatare biocompatible can in aqueous environment, provide oil free lubricity(Drobeck et al. Langmuir. 24(4):1484-8).

The surface modifications imparted by graft and block co-polymers hereindescribed may include, but are not limited to changes in wettability,steric modification or steric stabilization, steric repulsion, surfaceexclusion effects, high surface water retaining characteristics, chargeneutrality, and osmotic repulsion. These and other effects on thesurfaces are important processes that may convey clinically meaningfulbenefits to subjects and to the performance of medical devices. Thetissue-adhesive sections of a bi-functional co-polymer in theformulations used in the methods of the present invention may becationic, in which case the polymer adheres to the biological surface byelectrostatic attraction, or may be hydrophobic in which case thepolymer adheres to the biological surface by hydrophobic interaction, ormay involve a covalent complex. In some cases, the wettability of thebiological surface may result in whole or in part from modification of aliquid layer adjacent the surface, such as the mucous layer adjacent thecornea. In some cases the tissue surface modification may simply imparta lubricious protective coating over ophthalmic tissue or extraoculardevices. It may also alter the tribological properties of tissues andextraocular devices.

Aspects of the invention include copolymer coated nanoparticles whichcan encompass active agents, including drugs (small molecule, chemical,pharmaceutical, biologic) or allergens (viruses, bacteria, yeast,prions). The nanoparticles have exposed on their surface (i.e., arecoated with) a copolymer, where the copolymer has a positively charged,or hydrophobic, or covalent bonding moiety and a hydrophilic moiety.Nanoparticles are not of sufficient size to encompass cells.Nanoparticles are not microspheres. Other aspects of the inventioninclude coating drug particles or particles containing drug to increasetheir utility by increasing half-life and/or decreasing interaction withbiologic phenomenon such as an immune response. For intramuscularinjection, the nanoparticle size should be 1mm or smaller diameter; forinhalation, the nanoparticle size should be 10 microns or smaller indiameter; for IV injection, the nanoparticle size should be 1 micron orsmaller in diameter.

Aspects of the invention include copolymer coated nanoparticles whichcan encompass active agents, including drugs (small molecule, chemical,pharmaceutical, biologic) or allergens (viruses, bacteria, yeast,prions). The nanoparticles have exposed on their surface (i.e., arecoated with) a copolymer, where the copolymer has a positively charged,or hydrophobic, or covalent bonding moiety and a hydrophilic moiety.Nanoparticles are not of sufficient size to encompass cells.Nanoparticles are not microspheres. Other aspects of the inventioninclude coating drug particles or particles containing drug to increasetheir utility by increasing half-life and/or decreasing interaction withbiologic phenomenon such as an immune response.

Nanoparticles of the invention have an effective average particle sizeof less than about 1000 nm (i.e., 1 micron), less than about 900 nm,less than about 800 nm, less than about 700 nm, less than about 600 nm,less than about 500 nm, less than about 400 nm, less than about 300 nm,less than about 250 nm, less than about 200 nm, less than about 150 nm,less than about 100 nm, less than about 75 nm, or less than about 50 nm,as measured by light-scattering methods, microscopy, or otherappropriate methods. By “an effective average particle size of less thanabout 2000 nm” it is meant that at least 50% by weight of the activeagent particles have a particle size less than the effective average,i.e., less than about 2000 nm, 1900 nm, 1800 nm, etc., when measured bythe above-noted techniques. In other embodiments of the invention, atleast about 70%, at least about 90%, or at least about 95% of the activeagent particles have a particle size less than the effective average,i.e., less than about 2000 nm, 1900 nm, 1800 nm, etc. For intramuscularinjection, the nanoparticle size should be 1 mm or smaller diameter; forinhalation, the nanoparticle size should be 10 microns or smaller indiameter; for IV injection, the nanoparticle size should be 1 micron orsmaller in diameter.

Nanoparticles of the invention can be made using, for example, milling,homogenization, or precipitation techniques. Exemplary methods of makingnanoparticulate compositions are described in the '684 patent. Methodsof making nanoparticulate compositions are also described in U.S. Pat.No. 5,518,187 for “Method of Grinding Pharmaceutical Substances;” U.S.Pat. No. 5,718,388 for “Continuous Method of Grinding PharmaceuticalSubstances;” U.S. Pat. No. 5,862,999 for “Method of GrindingPharmaceutical Substances;” U.S. Pat. No. 5,665,331 for“Co-Microprecipitation of Nanoparticulate Pharmaceutical Agents withCrystal Growth Modifiers;” U.S. Pat. No. 5,662,883 for“Co-Microprecipitation of Nanoparticulate Pharmaceutical Agents withCrystal Growth Modifiers;” U.S. Pat. No. 5,560,932 for“Microprecipitation of Nanoparticulate Pharmaceutical Agents;” U.S. Pat.No. 5,543,133 for “Process of Preparing X-Ray Contrast CompositionsContaining Nanoparticles;” U.S. Pat. No. 5,534,270 for “Method ofPreparing Stable Drug Nanoparticles;” U.S. Pat. No. 5,510,118 for“Process of Preparing Therapeutic Compositions ContainingNanoparticles;” and U.S. Pat. No. 5,470,583 for “Method of PreparingNanoparticle Compositions Containing Charged Phospholipids to ReduceAggregation,” all of which are herein incorporated by reference.

The resultant nanoparticulate active agent compositions can be utilizedin solid or liquid dosage formulations, such as controlled releaseformulations, solid dose fast melt formulations, aerosol formulations,nasal formulations, lyophilized formulations, tablets, capsules, solidlozenge, powders, creams, ointments, etc.

The nanoparticles of the invention can be contacted with a copolymerhaving a positively charged, or hydrophobic or covalent bonding moietyand a hydrophilic moiety after attrition. One or more secondary surfacestabilizers may also be added before or after attrition. The activeagent particles can be reduced in size in the presence of a copolymerhaving a positively charged, or hydrophobic or covalent bonding moietyand a hydrophilic moiety. Other compounds, such as a diluent, can beadded to the copolymer active agent/surface stabilizer compositionduring the size reduction process (e.g., milling). Dispersions can bemanufactured continuously or in a batch mode.

Another method of forming the copolymer coated nanoparticles of theinvention is by microprecipitation. This is a method of preparing stabledispersions of copolymers or active agents in the presence of one ormore surface stabilizers and one or more colloid stability enhancingsurface active agents free of any trace toxic solvents or solubilizedheavy metal impurities. Such a method comprises, for example: (1)dissolving the copolymer or active agent in a suitable solvent; (2)adding the formulation from step (1) to a solution containing acopolymer having a positively charged, or hydrophobic or covalentbonding moiety and a hydrophilic moiety and/or active agent andoptionally one or more secondary surface stabilizers, to form a clearsolution; and (3) precipitating the formulation from step (2) using anappropriate non-solvent. The method can be followed by removal of anyformed salt, if present, by dialysis or diafiltration and concentrationof the dispersion by conventional means.

Exemplary homogenization methods of preparing copolymer coatednanoparticles are described in U.S. Pat. No. 5,510,118, for “Process ofPreparing Therapeutic Compositions Containing Nanoparticles.” Such amethod comprises dispersing nanoparticles in a liquid dispersion mediumin which the copolymer or active agent is soluble, followed bysubjecting the dispersion to homogenization to reduce the particle sizeof the active agent to the desired effective average particle size. Thenanoparticles can be reduced in size in the presence of a copolymerhaving a positively charged, or hydrophobic or covalent bonding moietyand a hydrophilic moiety and/or active agent, and, if desired, one ormore additional surface stabilizers. Alternatively, the nanoparticlescan be contacted with the copolymer and/or active agent and, if desired,one or more additional surface stabilizers either before or afterattrition. Other compounds, such as a diluent, can be added to theactive agent/copolymer composition either before, during, or after thesize reduction process. Dispersions can be manufactured continuously orin a batch mode.

The copolymer coated nanoparticles of the invention may contain anactive agent. The active agent may be contained within the nanoparticle,on the surface of the nanoparticle, in the copolymer coating of thenanoparticle, or on the surface of the copolymer coating of thenanoparticle. Active agents that can be used with the copolymer coatednanoparticle include without limitation drugs (small molecule, chemical,pharmaceutical, biologic) or allergens (viruses, bacteria, yeast,prions). Active agents may be therapeutic, or diagnostic agent. Atherapeutic agent can be a pharmaceutical agent, including biologicssuch as proteins, peptides, and nucleotides, or a diagnostic agent, suchas a contrast agent, including x-ray contrast agents. The active agentexists either as a discrete, crystalline phase, an amorphous phase, asemi-amorphous phase, a semi-crystalline phase, or mixtures thereof. Thecrystalline phase differs from a non-crystalline or amorphous phasewhich results from precipitation techniques, such as those described inEP Pat. No. 275,796. Two or more active agents can be used incombination.

The active agent can be selected from a variety of known classes ofdrugs, including, for example, proteins, peptides, nucleotides,anti-obesity drugs, nutraceuticals, dietary supplements, carotenoids,corticosteroids, elastase inhibitors, anti-fungals, oncology therapies,anti-emetics, analgesics, cardiovascular agents, anti-inflammatoryagents, anthelmintics, anti-arrhythmic agents, antibiotics (includingpenicillins), anticoagulants, antidepressants, antidiabetic agents,antiepileptics, antihistamines, antihypertensive agents, antimuscarinicagents, antimycobacterial agents, antineoplastic agents,immunosuppressants, antithyroid agents, antiviral agents, anxiolyticsedatives (hypnotics and neuroleptics), astringents, beta-adrenoceptorblocking agents, blood products and substitutes, cardiac inotropicagents, contrast media, corticosteroids, cough suppressants(expectorants and mucolytics), diagnostic agents, diagnostic imagingagents, diuretics, dopaminergics (antiparkinsonian agents),haemostatics, immunological agents, lipid regulating agents, musclerelaxants, parasympathomimetics, parathyroid calcitonin andbiphosphonates, prostaglandins, radio-pharmaceuticals, sex hormones(including steroids), anti-allergic agents, stimulants and anoretics,sympathomimetics, thyroid agents, vasodilators, and xanthines.

Exemplary nutraceuticals and dietary supplements are disclosed, forexample, in Roberts et al., Nutraceuticals: The Complete Encyclopedia ofSupplements, Herbs, Vitamins, and Healing Foods (American NutraceuticalAssociation, 2001), which is specifically incorporated by reference. Anutraceutical or dietary supplement, also known as phytochemicals orfunctional foods, is generally any one of a class of dietarysupplements, vitamins, minerals, herbs, or healing foods that havemedical or pharmaceutical effects on the body. Exemplary nutraceuticalsor dietary supplements include, but are not limited to, lutein, folicacid, fatty acids (e.g., DHA and ARA), fruit and vegetable extracts,vitamin and mineral supplements, phosphatidylserine, lipoic acid,melatonin, glucosamine/chondroitin, Aloe Vera, Guggul, glutamine, aminoacids (e.g., iso-leucine, leucine, lysine, methionine, phenylanine,threonine, tryptophan, and valine), green tea, lycopene, whole foods,food additives, herbs, phytonutrients, antioxidants, flavonoidconstituents of fruits, evening primrose oil, flax seeds, fish andmarine animal oils, and probiotics. Nutraceuticals and dietarysupplements also include bio-engineered foods genetically engineered tohave a desired property, also known as “pharmafoods.”

Active agents to be administered in an aerosol formulation arepreferably selected from the group consisting of proteins, peptide,bronchodilators, corticosteroids, elastase inhibitors, analgesics,anti-fungals, cystic-fibrosis therapies, asthma therapies, emphysematherapies, respiratory distress syndrome therapies, chronic bronchitistherapies, chronic obstructive pulmonary disease therapies,organ-transplant rejection therapies, therapies for tuberculosis andother infections of the lung, fungal infection therapies, respiratoryillness therapies associated with acquired immune deficiency syndrome,an oncology drug, an anti-emetic, an analgesic, and a cardiovascularagent.

A description of these classes of active agents and a listing of specieswithin each class can be found in Martindale, The Extra Pharmacopoeia,Twenty-ninth Edition (The Pharmaceutical Press, London, 1989),specifically incorporated by reference. The active agents arecommercially available and/or can be prepared by techniques known in theart.

Exemplary active agents to be applied to mucous include dentalapplications, such as oral bioadhesive nanoparticulate lidocainformulations, bioadhesive nanoparticulate fluoride treatments,application to the lungs, throat, GIT, application to wounds, etc. Alsoincluded is application to the throat using a liquid containing abioadhesive nanoparticulate formulation containing, for example, mentholor other numbing compound for treatment of coughs or sore throats. Thestomach and GIT can also be treated using bioadhesive formulations. Thisis particularly useful for treatment of diseases associated with themucous of the gastrointestinal tract, such as Crohn's Disease. Otherpharmaceutical therapeutic methodologies include oral dosing, nasaladministration, vaginal administration, ocular administration, colonic,and subcutaneous administration.

The compositions of the invention also encompass food products. Forexample, spice, oleoresin, flavor oil, color, or chemicals are oftenadded during food processing to produce the desirable flavors, taste,and appearance. These agents can be included in a bioadhesivenanoparticulate composition of the present invention for increasedadhesion to biological surfaces. Bioadhesive nanoparticulate flavoringagents could be used in products such as gums to produce prolongedflavor.

Other exemplary uses of the novel bioadhesive formulations are provided:teeth can be treated with teeth whiteners or fluoride bioadhesivecompositions; bones can be treated with calcium bioadhesivecompositions; nails can be treated with color or strengtheningbioadhesive formulations; insects or pests can be treated withinsecticides or other toxic compositions to the pest. In sum, thecompositions are useful in treating any biological surface, or a surfacederived from a biological material. Feathers and scales of animals canbe treated, as well as other animal biological surfaces such as chitin.

For the purpose of this invention, the description that follows below onthe different types of graft and block co-polymers applies to inventionembodiments of formulations for ophthalmic and medical topical andpharmaceutical use, and to the use of these graft and block co-polymersin the setting of extra- and intra-corneal ophthalmic devices ascoatings, and for ways of treating, storing, or manufacturing, orreapplying the coatings to these devices:

Examples of cationic polymer sections of graft or block co-polymers offormulations useful in the methods of the present invention include, butare not limited to: poly(L-lysine) (PLL), polylysine [includingpoly-D-lysine (PDL), poly-DL-lysine, poly-e-CBZ-D-lysine,poly-e-CBZ-DL-lysine, or poly-e-CBZ-L-lysine) polyornithine (i.e.,poly-DL-ornithine, poly-L-ornithine or poly-S-CBZ-DL-ornithine), and thelike)], [note Pub. No.: WO/1993/018649 International Application No.:PCT/US1993/002609 contains additional descriptions of polycationic watersoluble graft copolymers and is incorporated here by reference],poly(2-vinyl pyridine, poly(4-vinyl pyridine) and vinyl co-polymerscontaining those repeat units, and poly(aminoethyl methacrylate) homo-and co-polymers containing N,N dimethylaminoethylmethacrylate) repeatunits. In general, polymers containing acrylates and acrylamides canserve as cationic sections of the bi-functional co-polymers. Additionalexemplary cationic polymers include, but are not limited to,poly(trimethylammonioethyl methacrylate chloride),poly-(2-(dimethylamino) ethyl methacrylate) (pDMAEMA),poly-(2-(dimethylamino) ethyl methacrylateco-methacrylic acid)(pDMAEMA-co-MAA) and poly-(2-methyl-acrylic acid2-[(2-dimethylamino-ethyl)-methylamino]-ethyl ester) (pDAMA), and cationguar gum. Another cationic polymer section which can be used is chitosan(a co-polymer of glucosamine and N-acetyl glucosamine where 5-100% ofthe repeat units are glucosamine) and synthetic derivatives thereof. Theuse of amines and other positively charged amino acids bound topolymeric configurations to mimic the cationic polymeric activities sodescribed. In general, molecular structures that can impart a positiveexternal charge on a block or graft copolymer or a dendrimer areincluded in the invention.Examples of hydrophobic polymer sections of bi-functional co-polymers offormulations useful in the methods of the present invention include, butare not limited to, long-chain aliphatic hydrocarbons, polyethylene,poly(propylene oxide), polystyrene, poly(methylmethacrylate),poly(butylenes oxide), and the like. In general, molecular structuresthat can impart an exposed hydrophobic domain on a block or graftcopolymer are included in the invention.The hydrophilic section of the polymer may be non-ionic if the tissueadhering section is cationic, or non ionic or anionic if thetissue-adhering section is non-ionic (and hydrophobic). Examples ofnon-ionic hydrophilic polymer sections of formulations useful in methodsof the present invention include, but are not limited to,poly(ethyleneglycol) (PEG), poly(vinylalcohol),poly(vinylpyrrolidinone), dextrans and the like. Examples of anionichydrophilic polymer sections include homopolymers and co-polymerscontaining, for example, acrylic acid, methacrylic acid, itaconic acid,maleic acid, styrene sulfonic acid, carboxymethylcellulose,carboxyethylcellulose, succinylated chitosan, cellulose sulfate, and thelike. Additional exemplary hydrophilic polymers include, but are notlimited to, poly(dimethylamino)ethyl methacrylate and poly hydroxypropylmethacrylate (PHPMA).The adsorbing segment could also comprise a boronate group selected fromthe group consisting of phenylboronic acid (PBA), 2-carboxyethaneboronicacid, 1,2-carboxyethaneboronic acid,.beta.,.beta.′-dicarboxyethaneboronate,.beta.,.gamma.-dicarboxypropaneboronate, 2-nitro- and4-nitro-3-succinamidobenzene boronic acids,3-nitro-4-(6-aminohexylamido)phenyl boronic acid,{4-[(hexamethylenetetramine)methyl]phenyl}boronic acid,4-(N-methyl)carboxamidobenzene boronic acid,2-{[(4-boronphenyl)methyl]ethylammonio}ethyl and2-{[(4-boronphenyl)methyl]diethylammonio}-ethyl groups,succinyl-3-aminophenylboronic add, 6-aminocaproyl-3-aminophenylboronicacid, 3-(N-succinimidoxycarbonyl)aminophenylboronate,p-(.omega.-aminoethyl)phenylboronate, p-vinylbenzeneboronate,N-(3-dihydroxyborylphenyl)succinamic acid,N-(4-nitro-3-dihydroxyborylphenyl)succinamic acid,O-dimethylaminomethylbenzeneboronic acid, 4-carboxybenzeneboronic acid,4-(N-octyl)carboxamidobenzeneboronic acid,3-nitro-4-carboxybenzeneboronic acid, 2-nitro-4-carboxybenzeneboronicacid, 4-bromophenylboronate, p-vinylbenzene boronate,4-(.omega.-aminoethyl)phenylboronate, catechol[2-(diethylamino)carbonyl, 4-bromomethyl]phenyl boronate, and5-vinyl-2-dimethylaminomethylbenzeneboronic acid.Adhesion resistant segments is another way to describe an importantcomponent of the copolymer. Adhesion resistant moieties could include:polyalkylene oxides, mixed polyalkylene oxides, polysaccharides,polyvinyl alcohol, poly-N-vinyl pyrrolidone, noncationic polyacrylates,noncationic polymethacrylates, and mixtures and copolymers of theseconstituents.

In general, the chains and block and graft copolymer moieties may be anylength, size, or molecular weight, and may have any number of repeatrates of the polymer components. Imparting beneficial properties is themain variable that will determine specific size, weights, chain lengths,and repeat specifications for applications described herein. As anexample, PEG of different molecular weights can be grafted onto thepoly-(L-lysine) at different ratios of PLL:PEG, to optimize the polymerarchitecture for wettability, steric stabilization, and proteinresistance. Additionally, other factors, such as cost, processdevelopment requirements, manufacturing, and biologic tolerability willgo into making specific size, weight, chain length, and repeatcharacteristic specification decisions. For these polymers,polydispersity may be acceptable over both wide and narrow ranges, andproduct specifications will be made based on this characteristic aswell. However, this invention claims the use of all sizes, weights,chain lengths, and repeat characteristics of these graft and blockco-polymers for uses and methods described herein.

Graft co-polymers may have a cationic (or non-ionic hydrophobic)backbone made from a polymer chosen from the list, supra, andhydrophilic grafts, or have a hydrophilic backbone, and cationic (ornon-ionic hydrophobic) grafts chosen from the list, supra. For graftco-polymers, grafts may arise from every repeat unit in the backbone ormay be intermittently spaced along the backbone (with uniform or randomfrequency). For example, a useful polymer in formulations for use in themethods of the present invention is PLL-g-PEG where the backbone is thecationic polymer poly(L-lysine) and the grafts are made from thehydrophilic polymer poly(ethylene glycol). The PLL backbone may be from3 repeat units to several thousand repeat units long, and the PEG graftsmay be from 1 to several thousand repeat units long. The PEG grafts maybe attached to every PLL repeat unit, every other PLL repeat unit, everythird repeat unit or less frequent. In one embodiment, there is a PEGgraft on average at every third PLL repeat unit. Regarding PLL, the PLLchain length can be any length, and made from PLL derived fromfermentation processes or synthetic polymerization reactions. Themolecular weight of the PLL backbone can be in the range of severalthousand Daltons (3,000 to 5,000) to tens of thousands of Daltons 15,000to 30,000, or higher. The chain length can be as short as ten lysinegroups or as long as one to two hundred (or longer) lysine groups. Thepolyamino acid chain bonds may be epsilon or alpha based. By block orgraft co-polymers it is meant to describe the architecture of thepolymer.

An additional exemplary polymer for use in formulations used in themethods of the present invention is PLL-g-dextran. Similar detailedanalysis and variability descriptions (as in the paragraph above) can bemade for PLL-g-dextran, and for many other graft co-polymers claimed inthis invention. Those skilled in the art will identify methods to mimicthe behavior of the copolymers herein described using different monomersand moieties, and those methods and compositions are claimed.

Block co-polymers or dendrimers comprising at least one block or moietythat is cationic and at least one block or moiety that is water solubleand non-ionic are also useful in formulations for use in methods of thepresent invention. In one embodiment, the co-polymer comprises at leastone block which is hydrophobic and at least one block which is watersoluble and anionic, cationic or non-ionic.

Examples of water soluble non-ionic co-polymer blocks or moietiesinclude, but are not limited to, poly(ethylene glycol) (PEG), polyvinylalcohol (PVA), poly(hydroxyethyl methacrylate) (pHEMA),poly(acrylamide), poly (vinyl pyrrolidone) (PVP), poly(ethyl oxazoline)(PEOX), polysaccharides, and copolymers of any two or more thereof.

Examples of water soluble anionic co-polymer blocks or moieties include,but are not limited to, polyacrylic acid (PAA), polymethacrylic acid,poly(sodium styrene sulfonate), carboxylated cellulosics such ascarboxymethylcellulose (CMC), poly(itaconic acid), poly(maleic acid),poly(acrylamidopropanesulfonic acid), anionic natural gums, anioniccarbohydrates, carageenan, alginates and hyaluronic acid.

Examples of water soluble cationic co-polymer blocks include, but arenot limited to, polymers based on vinyl pyridine,N,N-dimethylaminoethylacrylate, N,N-dimethylaminoethylmethacrylate,other acrylate and acrylamide polymeric structures, allyl tri(alkyl)ammonium halides, poly(amino styrene), certain types of cation guargums, chitosan, polyethyleneimine, polyallylamine, polyetheramine,polyvinylpyridine, polysaccharides having a positively chargedfunctionality thereon, polyamino acids such as, but not limited to,poly-L-histidine, poly-im-benzyl-L-histidine, poly-D-lysine,poly-DL-lysine, poly-L-lysine, poly-ε-CBZ-D-lysine,poly-ε-CBZ-DL-lysine, poly-ε-CBZ-L-lysine, poly-DL-ornithine,poly-L-ornithine, poly-Δ-CBZ-DL-ornithine, poly-L-arginine,poly-DL-alanine-poly-L-lysine, poly(-L-histidine, L-glutamicacid)-poly-DL-alanine-poly-L-lysine, poly(L-phenylalanine, L-glutamicacid)-poly-DL-alanine-poly-L-lysine, and poly(L-tyrosine, L-glutamicacid)-poly-DL-alanine-poly-L-lysine, copolymers of L-arginine withtryptophan, tyrosine, or serine, copolymers of D-glutamic acid withD-lysine, copolymers of L-glutamic acid with lysine, ornithine, ormixtures of lysine and ornithine, and poly (L-glutamic acid), and theuse of amines and other positively charged amino acids bound topolymeric configurations to mimic the cationic polymeric activities sodescribed.

Examples of hydrophobic co-polymer blocks include, but are not limitedto, alkanes, alkenes, alkynes, poly(isobutylene), polyesters such aspoly(caprolactone) (PCL), poly(lactic acid) (PLA), poly(glycolic acid)(PGA), and copolymers therefrom (PLGA), polyamides such as nylon(6,6)and Nylon(12), polyurethanes, poly(propylene oxide), poly(tetramethyleneoxide), polyethylene, polypropylene, polystyrene, poly(acrylates) suchas polymethyl acrylate (PMA), poly(methacrylates) such aspoly(methylmethacrylate) (PMMA), poly(sulfones), poly(etheretherketones)(PEEKs), poly(phosphazines), poly(carbonates), poly(acetals) andpoly(siloxanes).

As described supra, a host of additional adsorbing agents can beutilized in the polymer construction.

As will be understood by the skilled artisan upon reading thisdisclosure, triblock configurations can be used. An exemplary blockco-polymer comprising a triblock configuration is PLURONIC® F127, alsoreferred to as Poloxamer 407, containing a poly(ethylene oxide)hydrophilic block (“PEO”), a poly(propylene oxide) hydrophobic block(“PPO”) and another PEO block. Other block co-polymers for use in thepresent invention may contain only one hydrophilic block and onehydrophobic block, or may contain several alternating blocks, forexample the PPO-PEO-PPO block co-polymers (PLURONIC®, block co-polymersbased on ethylene oxide and propylene oxide, BASF, Florham Park, N.J.).Additional exemplary PLURONIC block co-polymers useful in the presentinvention include, but are not limited to, PLURONIC 10R5, PLURONIC 17R2,PLURONIC 17R4, PLURONIC 25R2, PLURONIC 25R4, PLURONIC 31 R1, PLURONIC F108 Cast Solid Surfacta, PLURONIC F 108 Pastille, PLURONIC F 108 Prill,PLURONIC F 108NF Prill Polaxamer 338, PLURONIC F 127 Prill, PLURONIC F127 NF, PLURONIC F 127 NF 500 BHT Prill, PLURONIC F 127 NF PrillPoloxamer 407, PLURONIC F 38, PLURONIC F 38 Pastille, PLURONIC F 68,PLURONIC F 68 Pastille, PLURONIC F 68 LF Pastille, PLURONIC F 68 NFPrill Poloxamer 188, PLURONIC F 68 Prill, PLURONIC F 77, PLURONIC F 77Micropastille, PLURONIC F 87, PLURONIC F 87 NF Prill Poloxamer 237,PLURONIC F 87 Prill, PLURONIC F 88 Pastille, PLURONIC F 88 Prill,PLURONIC F 98, PLURONIC F 98 Prill, PLURONIC L 10, PLURONIC L 101,PLURONIC L 121, PLURONIC L 31, PLURONIC L 35, PLURONIC L 43, PLURONIC L44, PLURONIC L 44 NF Polaxamer 124, PLURONIC L 61, PLURONIC L 62,PLURONIC L 62 LF, PLURONIC L 62D, PLURONIC L 64, PLURONIC L 81, PLURONICL 92, PLURONIC L44 NF INH surfactant Polaxamer 124, PLURONIC N 3,PLURONIC P 103, PLURONIC P 104, PLURONIC P 105, PLURONIC P 123Surfactant, PLURONIC P 65, PLURONIC P 84, and PLURONIC P85. Whereapplicable, all particle sizes of the block co-polymers are included,for example PLURONIC F127 and PLURONIC F87 are available as prill andmicroprill products. Non-ionic surfactants, for example, containing ahydrophobic segment and a PEO block are considered here as blockco-polymers. The use of these commercially available agents specificallyin combination in formulations with bifunctional copolymers heredescribed including cationic and other adsorbing moieties coupled withhydrophilic or non reactive elements is identified.

Additional exemplary block or graft co-polymers or dendrimers which canbe used in the present invention are disclosed in U.S. Pat. Nos.5,578,442 and 5,834,556, as well as patents: U.S. Pat. Nos. ,462,990;5,627,233; 5,567,440; 5,849,839; 5,820,882; 5,380,536; 5,232,984;6,231,892; 6,743,521; 7,316,845; 2,286,590; 6,596,267; 7,029,688;6,350,527; 6,652,902. Teachings of each of which are herein incorporatedby reference in their entirety.

By “formulations” it is meant the particular mixture of base chemicalsand additives required for a product.

As mentioned previously, the examples of multi-functional orbifunctional copolymers described above can be used in formulations fortopical ophthalmic use and for coating extraocular and intra-cornealdevices. However, the invention is not limited only to those entitiesdescribed above, other variations or types of graft and blockco-polymers having properties described herein is an embodiment of theinvention. The invention is not limited only to those block and graftco-polymers named and described in detail herein.

The block or graft co-polymers are included in formulations for use inthe methods of the present invention at weight percent concentrationsranging between 0.001% and 40%, more typically 0.01% to 25%, of theformulation. An effective amount is claimed. In the Examples in whichthe formulation is described at present, the amounts of co-polymers arebetween about 0.1% and 2%. In addition the amount of bi-functionalco-polymer can fall within other smaller ranges e.g., 0.01% to 3%, 0.1%to 2.5%, or 0.5 to 2%.

The combinations of different copolymers can be that each is(weight/weight) between 0.01% and 2% but can have ranges high as wide as0.01% and 5%. An effective amount is an amount that increases thewettability of a given biological surface, or contact lens surface; orthat changes or modifies the surface in a favorable way; suchmodification can include but is not limited to steric stabilization,steric repulsion, high surface water retaining capacity, chargeneutrality, surface exclusion effects, or osmotic repulsion. The extentof increase in wettability will vary with the application and with thedisease or disease symptom that is being treated or prevented. Aneffective amount is an amount that leads to beneficial biologicalsurface modification characteristics, as described above.

The bi-functional copolymers are used in any of these concentrations toimpart favorable characteristics to a product for use in humans or othermammals either with or without an active pharmaceutical ingredient. Anactive pharmaceutical ingredient can be added to these formulations andthe active agent will be used in weight percent concentrations frombetween 0.0001% to 40%, more typically from 0.01% to 10%. The copolymerscan be used with a preservatives such as, but are not limited to,Polixetonium, polyquaternium-42, Polyquaternium-1, Polyquat,Alkyl-hydroxy benzoate preservatives, parabens, hydrogen peroxide,benzalkonium chloride, cetylpyidimine chloride, cetalkonium chloride,sodium perborate, Purite, disappearing preservatives, Polyhexamethylenebiguanide (PHMB), chlorobutanol, Benzododecinium bromide, “Ionicbuffered system”, povidone, silver, silver sulfate, betadine, and otherantiseptics and proprietary and non-proprietary preservatives. Thepolymers herein described can be used in combination with surfactants oremulsifier. Surfactant means a chemical agent capable of reducing thesurface tension of a liquid in which it is dissolved. Emulsifier means asubstance which stabilizes an emulsion by increasing its kineticstability.

An important aspect of this invention is formulations combiningbi-functional copolymers with one or more preservatives, surfactants,and/or emulsifiers.

Polydispersity in these graft and block co-polymers can be eitherminimal or there can be significant amounts of polydispersity, and theformulations can still be effective.

Viscosity can important in these formulations. Typically, with currenttechnology, higher viscosity eye drop formulations lead to longersurface changes on an eye but are associated with visual blur anddissatisfaction by the user. There are advantages to lower viscosityagents, and in certain formulations described herein a lower viscositycan be beneficial as there will be less visual blur imparted when an eyedrop is applied. In the current embodiment, the viscosity isapproximately 2.7 centipoise (cP), however, it can be higher or lower.2.7 cP is lower than the viscosity of most commercially availableartificial tears. In some embodiments a higher viscosity formulation maybe preferred, however. Viscosity can be adjusted by adjustingconcentrations of the graft and/or block co-polymers, or by adjustingother excipients to affect viscosity. Useful viscosities are in therange of 1 cP up to and including 100 cP or higher, but typically theviscosity would be between 2 and 30 cP.

Additional exemplary components which can also be incorporated intopharmaceutical formulations and coatings for use in the presentinvention include, but are not limited to PLURONIC gelling agents suchas, but not limited to F127, F108 as well as additional PLURONIC agentslisted supra. Furthermore, in one embodiment, these components are usedat fractions below that required for gelling activity.

Other components (either active or inactive ingredients) which can beincluded in these pharmaceutical formulations include, but are notlimited to, lipids, oils, surfactants, water, lubricating polymers,typical surfactants, buffers, salts, physiologic ions, proteins, topicalemollients, excipients typically used in oral, topical, mucosal,dermatologic and ophthalmic formulations, lubricants such as PEG 400,carboxymethylcellose, hydroxypropyl methylcellulose, mineral oil,propylene glycol, glycerin, hypromellose, white petrolatum, polyvinylalcohol, liposomes, mannitol, hydroxypropyl guar, dextran 70,viscoelastics, guar gum, alginates, and hyaluronic acid, as well ascombinations thereof. Specifically, the use of the herein describedcopolymers for ophthalmic indications in combination with active agentsdescribed in the OTC Monograph 21CFR349.14 is claimed. Additionalingredients may include those routinely included in shampoos, soaps, andconditioners. Such components may be included in the formulations invarying percentages ranging from less than 0.1% to 99% w/w %, morepreferably less than 1% to 10% w/w %. These are wt %'s relative to thetotal formulation.

Other components which can be included in these pharmaceuticalformulations include preservatives such as, but are not limited to,Polixetonium, polyquaternium-42, Polyquaternium-1, Polyquat,Alkyl-hydroxy benzoate preservatives, parabens, hydrogen peroxide,benzalkonium chloride, cetylpyidimine chloride, cetalkonium chloride,sodium perborate, Purite, disappearing preservatives, Polyhexamethylenebiguanide (PHMB), chlorobutanol, Benzododecinium bromide, “Ionicbuffered system”, povidone, silver, silver sulfate, betadine, and otherantiseptics and proprietary and non-proprietary preservatives. Also,PLL-g-PEG, or other cationic components of graft or block co-polymersmay act as a preservative. Included in this invention, also, is the useof cationic moieties in the graft or block co-polymers to serve as apreservative for a formulation containing these agents. Poly (L) lysine,alone, not coupled with a second polymer, as an ophthalmic preservative,for example, is included as an invention in this filing.

Further, in some embodiments, the formulations and coatings may includeone or more additional active pharmaceutical ingredients. Examplesinclude, but are in no way limited to anesthetics, antibiotics,anti-virals, anti-inflammatory agents, intraocular pressure loweringagents, artificial tears, lubricating products, dilating agents,immunosuppressives, anti-angiogenic agents, proteins, peptides,neuroprotectants, small molecules, growth factors and antibodies. Byactive pharmaceutical ingredient or API it is also meant to be inclusiveof genes and/or gene transfer agents such as adenovirus, AAD and nonviral vectors for which transfer and transfection is enhanced viaformulations of the present invention. Formulations may be used inaccordance with the present invention to deliver an activepharmaceutical ingredient which acts locally at the surface or in thetissue to which it is delivered. Alternatively, or in addition,formulations may be used in accordance with the present invention todeliver an active pharmaceutical ingredient which is then absorbedthrough the tissue and has a systemic or distal effect. In someembodiments, delivery of the active pharmaceutical ingredient isfacilitated by external energy such as, but not limited to,iontophoresis, sonic energy, heat, microneedles, micropore creatingdevices such as lasers or high pressure gas.

Pharmaceutical formulations are a composition suitable for internal,topical, or ocular administration to an animal, including humans. Thecarrier is a pharmaceutically acceptable excipient in which theco-polymer is admixed.

“Eye lubricant” can be defined as the API included in artificial tearproducts based on the FDA OTC monograph 21 CFR 349.14 for ophthalmics.

pH of formulations of the present invention is in a physiologic rangedepending upon the site of administration and the site of the biologicalsurface or membrane that is to be modified. Typically, the pH is above3, e.g., above 5.6 and below 9. The pH is preferably between 6.5 and 8.Hypertonic and hypotonic formulations are claimed. Water is well knownas an important excipient in topical ophthalmic formulations.

The ability of a pharmaceutical formulation comprising a graft or blockco-polymer with a positively charged or hydrophobic moiety and ahydrophilic moiety to change wettability and adhere to and lubricate andmodify a biological surface was demonstrated. Thus, the tribologicalproperties of the surface was modified. Importantly, these formulationsmay have very low viscosity (lower than most artificial tear products onthe market). For example, the formulation tested in humans had aviscosity of 2.7 cP. Thus, the results are not due to a simplemechanical thickening of the tear film.

The ability of formulations comprising either the graft co-polymerPLL-g-PEG or the block co-polymer F127 to change wettability wasdemonstrated in vitro via contact angle experiments, as described inExample 2. FIG. 1 and FIG. 2 demonstrate the ability of exemplaryformulations comprising either 1, 0.5 or 0.1% weight/weight PEG-PLL or1% F87 or F127 weight/weight, respectively, to decrease the contactangle of water as compared to a control of deionized water. The abilityof these formulations to decrease contact angle is indicative of theirability to adhere to negatively charged surfaces such as biologicalsurfaces and/or membranes and change wettability of the biologicalsurface and/or membrane. Through the understanding of one skilled in theart, these formulations will thus also impart surface modifications suchas steric stabilization on the tissue to which the bi-functional copolymers adhere.

Important to the management of ocular surface conditions is an approachto protect the cornea and bulbar conjunctiva. These parts of the eye areexposed to the environment when the eye is open. When the eye is open, apatient with an unstable tear film will develop symptoms such asburning, irritation, and/or the sensation of dryness. Reflexive tearingmay or may not be present. Eventually, dry eye syndrome can develop anda host of signs and symptoms are associated with dry eye. One aspect todry eye is the evaporation rate of the tears. Faster evaporation,physically, from a surface leads to cooling of that surface. Typically,it is the lipid layer, or lipid components, of the tear film that resistevaporation. The lipids are part of a liquid structure (such as oils)that is typically more resistant to evaporation than water basedsolutions, especially low viscosity solutions.

There exists a host of conditions for which heat therapy is recommendedand effective. Heat can be used to treat eyelid conditions, meibomiangland dysfunction, lacrimal drainage conditions, chalazions, styes,infections, and other eye conditions. Heat is also used to treat skinand mucous membrane conditions such as infections, abscesses, andinflammatory conditions.

Therefore, there is a need in the art for a multi-functional polymerbased solution that can maintain or increase ocular surface temperatureor the surface temperature of the skin and other mucous membranes.Disclosed herein are multi-functional graft or block copolymer basedsolutions that are effective in methods for increasing surfacetemperature, particularly ocular surface temperature. Thesemulti-functional graft or block copolymer based solutions may encompassmany forms, such as a formulation for an artificial tear, (especially alow viscosity solution), or a topical ophthalmic polymer-includingproduct including those that can be used with contact lenses or in theformulations of drug products for the eye, that could maintain orincrease ocular surface temperature.

A polymer based solution comprising the multi-functional graft or blockcopolymer disclosed herein generates an increase in ocular surfacetemperature immediately after instillation as well as for prolongedperiods such as 15 or even 30 minutes following instillation, andprovides an opportunity for clinical treatment success in patients atrisk of or suffering from tear film abnormalities may exist with such aformulation. There are a more than one patient subpopulations that wouldbenefit from increasing ocular surface temperature. For example,patients with blepharitis, patients who have ocular discomfort, patientswho are status post eye surgery, patients with ocular allergy, patientswith ocular inflammation, and patients with or at risk for ocularinfections. It is important to focus on blepharitis patients in thisapplication. Blepharitis is inflammation of the eye lid margin.Blepharitis is a common cause of ocular irritation and discomfort in allage groups and ethnicities. There remains important unmet need in thetreatment of blepharitis. This condition can lead to permanent changesof the eyelid margin or vision change from corneal problems. Theinflammation affects the eyelid itself, and often secondarily thecorneal surface as well as tear quality. Currently, a recommendedtreatment is warm compresses to the eyelids and the eyelid margin. Theeyelash margin and areas anatomical close to there are where theMeibomian glands secrete their lipid component of tears. The lipids inthese secretions can become more viscous, less liquid-like, and evensolid (like many oils and fats) at lower temperatures. A treatment forMeibomian gland disease is using a formulation containing amultifunctional graft copolymer on the eye. The topical formulation willnecessarily bathe the eyelid margin as well. Note, the tear lake, orlacrimal lake, which is essentially the volume of tears in an eye, is 7to 10 microliters. The maximum that the tear lake can hold is about 20to 30 microliters after which tears leave the tear lake and wet theeyelids (tearing). The volume of an eye drop is about 40 microliters. Itis obvious that when an eye drop is applied to the eye, excess tearswill wet the eyelid. This approach is one way that the lid margin ormeibomian gland surface will be wetted with the topical polymerformulation when instilled as an eye drop. Additionally, however, theeyelid margin is generally directly opposed to the surface of the eye.Indeed, the opening and closing of the eyelid force the excess tearsnasally through the lacrimal drainage system. Thus, the lid marginitself, and the meibomian glands contained within will be coated withtears. The lid margin is wetted from even a smaller volume of topicalformulation—tearing or instillation of volumes that would cause themaximum volume of the tear lake to be exceeded are not necessary to wetthe lid margin and meibomian glands. In one embodiment of the presentinvention the Meibomian glands and/or the lid margin have the restingtemperature raised by the instillation of a multifunctional graftcopolymer. There will be clinical benefits for those suffering frommeibomian gland dysfunction and/or blepharitis by raising thetemperature of the lid margin. This approach is a novel and differentapproach than what is used clinically which includes warm compresses,and even a device called “Lipiflow” heats the eyelids to treatblepharitis. This novel approach of using a topically appliedmultifunctional graft copolymer to increase the lid margin temperatureshould have great clinical utility.

Furthermore, the palpebral conjunctiva also directly contacts thesurface of the eye. Since the Meibomian glands reside within the eyelid, there is also the advantage with this approach of placing themultifunctional graft copolymer in the eye irrespective of excesstearing to coat the lid margin. Since the ocular surface is higher,direct heat transmission will take place as a transfer of heat from theocular surface to the palpebral conjunctiva and the eyelids themselves.A new treatment for blepharitis is herein described. A new treatment toenhance meibomian gland secretions or the lipid component of tears, isthus described. A new treatment for Meibomian gland dysfunction isdescribed. This new treatment for eyelid margin disease will alsobenefit mattering of the eyelid as the build up around the lashes willbe more easily reduced with a change in temperature at the eyelidmargin. Patients with blepharitis can be treated with a form of eattherapy through the application of a multifunctional graft copolymer tothe eye. Note, multifunctional graft copolymer refers to PLL-g-PEG andother graft copolymer molecules with a cationic backbone and hydrophilicside chains. There is enough novelty with this approach such that awider range of graft copolymers is also included as a novel embodimentof this discovery.

The definition of an increase in temperature is at least 0.1 degreesCelsius. In some embodiments, and increase of 0.5 degrees Celsius ispossible. An increase of 1 degree Celsius is also possible. Greaterincreases in temperature may be seen with this technology. Temperatureis a continuous variable, and viscosity of lipids and oils vary withtemperature. Thus, any increase in temperature, will decrease lipidviscosity and help lipid tear flow into the eye. For the purposes ofbeing measurable and clinically significant, an increase in ocularsurface temperature, or the temperature of a mucous membrane (eyelidmargin included) is 0.1 degree Celsius. Likewise, heat is a therapy forcertain conditions. Since temperature is a continuous variable, anyincrease in temperature to target tissue should have a beneficialeffect. Increasing the eyelid temperature by 0.1 degree Celsius isconsidered a temperature increase. Such an increase (and greater) hasbeen observed directly with the topical application of a multifunctionalgraft copolymer.

Other patients that may benefit from this technology and variousembodiments of this invention include those with bacterial infections ofthe eye, conjunctiva, or eyelids, plus the skin and other mucousmembranes. Patients with corneal infections or viral keratitis orconjunctivitis will benefit from this treatment. Patients with acondition where increasing the temperature of the tissue is felt to betherapeutic will benefit from the use of these multifunctional graftcopolymers. Patients with dry skin and dry mouth will benefit. Patientswith non-healing ulcers whether due to vascular compromise or other willbenefit as heat causes vasodilation and increases blood flow to aregion. The use of these multifunctional graft copolymers can likewisebe used to prevent infection because increased blood flow can help thebody naturally prevent infection in certain circumstances. The treatmentfor a stye or chalazion involves heat. Thus, another embodiment is theapplication of the graft multifunctional copolymer to the eye and eyelidis a treatment for stye and chalazion. Patients with orbital orpreseptal cellulitis can be treated with these formulations. Otherpatients that may benefit from this method of increasing the temperatureof skin or mucous membranes by contacting the tissue with amultifunctional graft copolymer include dry eye patients to enhancelipid flow, glaucoma patients to help reduce the risk of bleb associatedendophthalmitis, corneal transplant subjects to help reduce the risk ofgraft infection, keratoprosthetic patients to help reduce the risk ofinfection and to enhance healing at the host/graft/device margin,post-operative pterygium surgeries where a conjunctival graft was used,limbal graft surgeries, post op cataract surgical patients where theincreased heat will enhance wound closure, arthritis patients, soremuscles, sore eyes, post-operative oculoplastic patients who haveundergone eyelid surgery, post-operative plastic surgery patients,patients with peripheral vascular disease, patients with infectious skinproblems, patients with an abscess, other signs of infection such asinduration, erythema, and pain, and patients with select auto-immuneskin conditions. Blepharitis and infectious conditions of the eyelid andthe eyelid margin remain a key opportunity for this new therapy. Mucousmembrane inflammation and infection remain a key treatment for othernon-ocular indications. The mucous membranes of the nares can be treatedwith multifunctional graft copolymers to enhance the prevention,treatment and clearance of viruses including the common cold andinfluenza. Elevated temperature helps certain types of immune cells workbetter, so there is a broad potential for benefit with this newtreatment for certain infectious conditions.

Toward that end the working examples demonstrate the effect on ocularsurface temperature of the multifunctional graft co-polymer in a topicalophthalmic formulation in a clinical setting. See Example 8 in which thesurface temperature after instillation was compared to the pre-exposureor pre-instillation ocular surface temperature.

Ocular surface temperature measurement can also be referred to as ocularthermography or ocular thermometry. Contact methods using thermistorscan be used to measure ocular surface temperature. Infrared imaging isthe preferred methodology. Thermal cameras also are used to measureocular surface temperature. One camera that specifically is useful hereis the Thermovision A40 from FLIR system Inc, which has a sensitivity todetect temperature change of 0.08 C, a frame rate 30 Hz, and imageresolution of 320×240 pixels across full field view and emissivity setat 0.98. Cameras that have a detection sensitivity of less than 0.1degree are preferred. The detection, however, should be able toaccurately identify changes of less than 1 degree Celsius. Any camerathat meets these specifications can in some embodiments be used to testfor the change in surface temperature. Other systems that can identifythese changes in ocular surface temperature include: thermography,infrared thermal imaging, dry eye, temperature distribution assessments,dynamic thermography, and ocular surface temperature assessments.

Similar approaches for temperature measurement can be used for otherbiological surfaces. For example, a multi-functional polymer basedsolution disclosed herein that can maintain or increase the surfacetemperature of the skin is helpful for treating skin conditions such aswith skin grafts and certain dermatological conditions that are slow toheal and are exposed to lengthy periods of cooling and dryingenvironments. This treatment with topically applied multifunctionalblock copolymers to raise skin temperature may benefit non-healingulcers, abscesses, induration, scar tissue prevention, and bacterialskin infections.

For methods of treating ocular or non-ocular skin grafts or other oculardisorders (such as eyelid infections and blepharitis) which endurecooling and slow healing conditions, a beneficial effect can be attainedby increasing the temperature of the ocular surface through the use of amulti-functional graft or block copolymer based solutions disclosedherein, A thermogram can report the ocular surface temperaturemeasurements (an ocular thermogram for the eye). Infrared cameras andsensors are optimally utilized for measuring ocular surface temperature(or biological surface temperature) in these situations.

It was unanticipated, surprising and important that the novel andproprietary topical formulation comprising a multi-functional polymerbased solution disclosed herein actually increased the ocular surfacetemperature, and that the effect of increasing the ocular temperaturegrew over time, especially in the setting of the subject holding theireyes open for prolonged periods.

Thus, it is a surprising discovery that a characteristic property of themulti-functional graft copolymer described herein is its ability toincrease ocular surface temperature when applied. Furthermore, when themulti-functional graft copolymer described herein is combined withlipids in an artificial tear formulation or other topical formulations,the ocular surface temperature raising characteristic remains.

Thus, encompassed herein are topical lipid containing ophthalmicformulations to which are added the multi-functional graft copolymerdescribed herein, methods of making said ophthalmic solutions, andmethods of using said ophthalmic solutions for the treatment ofophthalmic diseases, disorders and conditions.

Not only can the formulation containing the multi-functional graftcopolymer described herein be delivered to the eye directly byinstillation, but a contact lens can serve as a vehicle for deliveringthis multifunctional graft co-polymer. In one embodiment the contactlens can be manufactured with the multi-functional graft copolymerdescribed herein. In another embodiment or the contact lens can besoaked after manufacture with the multi-functional graft copolymerdescribed herein. In either embodiment, there will be somemulti-functional graft copolymer described herein that leaves thecontact lens either from the solution coating it or from the lenspolymer directly via some elution.

Thus, described herein is a method to increase the ocular surfacetemperature of the contact lens containing the multi-functional graftcopolymer described herein in wearers of said contact lens as well asincreasing the temperature of the eye's ocular surface. Furthermore, bywearing a contact lens coated with containing or exposed to aformulation with a multifunctional graft copolymer, the prescribingphysician and patient can treat an underlying condition, such asblepharitis, chalazion, stye, or corneal abrasion, to help improve thecondition. See example (Example 9.). Furthermore, the increasedtemperature of the eye's ocular surface will remain in place in contactlens wearers because the ocular surface temperature can also be measuredat the conjunctiva, which is not covered by a contact lens. Thus, thetemperature will be higher at the contact lens itself, and on theeyeball. The effect is demonstrated as compared to an untreated lens onthe eye of a subject. The presence of said multi-functional polymers andtheir variations as discussed in detail elsewhere herein exert theireffect on the ocular surface temperature when the temperature iscompared to a pre-instillation or pre-exposure baseline.

Importantly, the instantly disclosed method of increasing thetemperature of the eye's ocular surface comprising contacting saidsurface with formulations comprising the graft copolymers and blockcopolymers disclosed herein is not limited to the ocular surface. Aformulation for topical use containing said graft copolymers and blockcopolymers decreases rates of evaporation from a biological surface suchas the exposed skin or mucosal surface of a mammal, including but notlimited to humans. Included biological surfaces are the ocular surface,nasal mucosa, oral mucosa (for example during mouth breathing), externalear, scalp, lips, eyelids and skin, as well as the bulbar conjunctivaand cornea. Further biological surfaces include surface of body partsthat find exposure to the external environment following injury orsurgery including but not limited to as open wounds, skin grafts, andresections, as they will benefit from the temperature increasing effectof said formulations comprising the graft copolymers and blockcopolymers disclosed herein to reduce evaporation and subsequent tissuecooling and drying. The range of final concentrations of formulationswith a multifunctional block copolymer are from 0.001% to 50%. Becauseof the surface adhering characteristics quite low formulations have aneffect. Higher strength formulations are a potential way to furtherincrease the effect or to provide longer lasting products.

Thus, a biological surface includes the ocular surface, the skin, andmucosal surfaces. Wounds and organ or tissue surfaces are also includedin the definition of biological surfaces. For the purposes of thisapplication, the relevance is highest when these biological surfaceshave exposure to an environment whereby there will be or there is thepotential for cooling of that surface when wet by evaporation. A liquid(generally water-based) product to be applied to such surfaces willnecessarily show cooling on that surface. By including the inventiondescribed herein, embodiments of formulations comprising themultifunctional graft copolymers in the solution applied to thebiological surface will show less cooling at the contact of saidsurface.

Thus the phrase “increase in surface temperature” not only comprises anincrease in surface temperature of up to 0.2, 0.4, 0.5, 0.6, 0.8, 0.9,1.0, 1.2, 1.4, 1.5, 1.6, 1.8, 2.0, 2.2, 2.4, 2.5, 2.6, 2.8, 3.0, 3.2,3.4, 3.5, 3.6, 3.8, 4.0, 4.2, 4.4, 4.5, 4.6, 4.8, 5 degrees F. or more,or an increase in surface temperature of up to 0.2, 0.4, 0.5, 0.6, 0.8,0.9, 1.0, 1.2, 1.4, 1.5, 1.6, 1.8, 2.0, 2.2, 2.4, 2.5, 2.6, 2.8, 3.0,3.2, 3.4, 3.5, 3.6, 3.8, 4.0, 4.2, 4.4, 4.5, 4.6, 4.8, 5 degrees C. ormore, with respect to the surface temperature immediately beforeapplication of said graft copolymers and block copolymers disclosedherein, but the phrase also encompasses a decrease in the amount ofcooling of the biological surface upon contact with said formulationscomprising the graft copolymers and block copolymers disclosed hereincompared to said formulations without the graft copolymers and blockcopolymers disclosed herein, i.e., a baseline of cooling, is part of theinvention. The comparison in these situations is the evaporation rate,or surface temperature, when a formulation without the copolymer isapplied versus a formulation with the copolymer. The minimal relevantchange, or improvement, is 0.1 degree Celsius. The highest change to beseen will 5 degrees Celsius or less.

An increase in surface temperature is a change of at least 0.1 degreeFahrenheit from the pre-contact surface temperature to the post-contactsurface temperature. Copolymer concentrations can range from 0.01% to50% in pharmaceutical compositions in the embodied methods. Preferredembodiments of copolymer concentrations are in the range of 0.01% to 3%.The multiple varieties of this structural effect—which is aphysicochemical finding or characteristic of the polymer—will be seenregardless of the specific design or composition of a multi-functionalpolymer with positive charges on one moiety and hydrophilic charges onthe other. The preferred embodiment is a multi-functional graftcopolymer with a positively charged backbone and hydrophilic sidechains. The same benefits can be obtained when formulating with otheractives (an active is an agent approved allowed by the FDA or otherregulatory body as a treatment for a specific indication), for exampleglaucoma, anti-inflammatory, anti-allergy, and anti-infective activesfor treatment of patients having glaucoma, allergies, an ocularinfection, including a microbial infection, a bacterial infection, aviral infection, a parasite infection and a fungal infection. Otherconditions have been mentioned elsewhere. See Example 10. Thus, thedecreased temperature effect upon contact with a biological surface holdtrue for any of the variations of multi-functional graft copolymers thatmimic the structure and physicochemical properties described herein.

Example 11. Shows the utility of using the multi-functional graftco-polymer in drug products for the eye with other actives such as thosefor treating glaucoma. The effect of the actives on intraocular pressurewere maintained in the presence of multi-functional graft copolymers,and the tear film was significantly stabilized. The method for addingthe polymer simply to a topical ophthalmic solution will have value incertain clinical situations. For example, it may be beneficial for aphysician to write an order wherein polymer is added to an alreadymanufactured formulation. The manufacturing process, as seen, can bevery simple. In one embodiment, the manufacturing process comprisessterile filtration of the formulation and simply adding the polymer toform a formulation is safe and effective. Autoclave formulation isanother approach to product sterilization.

A “meaningful baseline” as mentioned in the claims regarding biologicalsurface temperature can be considered or is for the purposes of thisapplication, any one of, but not limited to: A) the temperature of thebiological surface at baseline if it is a mucous membrane, or B) thetemperature of the ocular surface prior to any eye drops instilled, orC) the temperature of the biological surface after it is coated orcontacted with a solution identical to the copolymer solution only thereis no copolymer component added, or D) the actual temperature of thebiological surface in some cases, or E) the temperature of thebiological surface when it is wetted with water in a smooth thin layer,or F) the temperature of the cornea, or G) the temperature of the bulbarconjunctiva.

As described in Example 3, Ex vivo experiments in porcine eyes alsoshowed a delay in pre-corneal water evaporation rates of approximately 2seconds longer compared to a control of Systane® lubricant eye drops(Active Ingredients: Polyethylene glycol 400 0.4% and propylene glycol0.3%. Inactive Ingredients: boric acid, calcium chloride, hydroxypropylguar, magnesium chloride, potassium chloride, purified water, sodiumchloride, zinc chloride.

As described in Example 1, experiments in a rabbit model also showedlack of toxicity, in other words, safety, for a formulation comprising1% glycerin with 1% PLL-g-PEG and 1% F127 as delivery of suchformulation of the present invention to a sensitive mucosal surfaceshowed no visible irritation in the Draize test and no histopathologicalchanges (14 day histopathology study).

Furthermore, as described in Examples 4 and 5, performance of theseblock and graft copolymer formulations was assessed in the eyes of humanvolunteers. Initial informal tolerability studies showed the eye dropwas well tolerated. For example, of the multiple initial humanexposures, there was no irritation or discomfort seen or reported in anysubject. Additionally, there were no reports of blur followinginstillation of 50 microliters or less. Actually, the eye drop wasdescribed as soothing on several occasions. As a further safety step,one volunteer received 1 milliliter of solution in the eye, multipletimes, and there was no irritation or discomfort. The ability to treatdry eye was then assessed using tear film break up time (TFBUT). TFBUTis indicative of the rate of evaporation of tears from the surface ofthe eye. The longer the TFBUT, the wetter the eye between blinks and theless likely a patient will have dry eye signs and symptoms. The firstassessments were informal and performed in only a couple of subjects,then a larger masked, randomized controlled study was carried out.Administration of a formulation comprising 1% glycerin with 1% PLL-g-PEGand 1% F127 resulted in prolongation of TFBUT. In the first informalstudies, the TFBUT was measured using fluorescein staining. The TFBUTwas prolonged versus active comparator on one occasion at early timepoints, and that increase was maintained for three hours. Subjectiveobservation of the tear film break up into smaller patches was confirmedvia a non-invasive wavescan of the tear surface. The larger, randomizedcontrolled trial studied three groups of patients (none/mild dry eye,mild/moderate dry eye, and moderate/severe dry eye). The total number ofsubjects was 18. Sixteen were analyzed with a noninvasive TFBUTassessment tool called the Tearscope™. In this study, there wasstatistically significant prolongation of the TFBUT at the 15 minutetime point and the two hour time point versus active comparator (aleading OTC dry eye artificial tear). The two additional subjectsunderwent wavescan assessments as an alternative to the Tearscope Plus™;the results in these two subjects also showed prolongation of the tearfilm compared to control. Accordingly, one embodiment of the presentinvention relates to use of these pharmaceutical formulations aslubricating opthalmic eye drops for treatment of, for example, dry eyesyndrome and contact lens intolerance. As will be understood by theskilled artisan upon reading this disclosure, however, alternativeophthalmic delivery means including, but not limited to, intraocular,periocular, conjunctival, subconjunctival, transconjunctival,peribulbar, retrobulbar, subtenons, transscleral, topical gel, topicaldispersion, intraorbital, intrascleral, intravitreal, subretinal,transretinal, choroidal, uveal, intracameral, transcorneal,intracorneal, intralenticular (including phakia and psuedophakia), andin or adjacent to the optic nerve, can be used. The invention can beused with polymeric and other devices for prolonged ophthalmic drugdelivery. The invention can be used with depot formulation to ease thetolerance of the eye to prolonged drug exposure.

Dry eye syndrome is a common and irritating problem for approximately 60million Americans. Five million Americans suffer from an advanced formof this condition. Dry eye syndrome can be due to a deficiency of theaqueous, lipid, or mucin component of tears. Dry eye syndrome is alsorelated to inflammation on the surface of the eye and irregularities ofthe tear film components. Aging and hormonal changes can play a role.Environment, as well as contact lenses and refractive surgery, caninitiate and/or exacerbate the problem.

Dry eye is treated with eye drops, punctual occlusion, and occasionallysystemically administered medicine, although eye drops are the mainstayof treatment. While most eye drop treatments are availableover-the-counter, there are two prescription eye drops inclusive ofsteroids and cyclosporine that help with dry eye. Most over-the-counterdry eye syndrome products are washed out through the lacrimal drainagesystem quite quickly thus rendering relief minimal. For example, in atleast two studies by Alcon, Systane had a beneficial effect on tear filmbreak up time that lasted up to 30 minutes. These studies demonstrateSystane™ is one of the longest acting over-the-counter dry eye products.The PLL-g-PEG formulation shows results improving tear film break uptime beyond two hours.

Accordingly, there is need for formulations of the present invention fortreatment of this condition.

Formulations can also be used in accordance with the present inventionto provide for prolonged coating of the ocular surface via themultifunctional properties of the graft and/or block co-polymers.Without being bound to any particular theory, it is believed that thehydrophilic domain(s) of the block or graft co-polymer, which keep theeye lubricated and help to retain tears on the surface, are anchored atthe surface of the eye (cornea and conjunctiva)for an extended period oftime through primarily interaction of positively charged domain(s) onthe block, graft, or backbone of the graft copolymer and secondarilythrough hydrophobic domains on the block, graft or backbone of thepolymer with the surface of the eye through either electrostaticattraction with the negatively charged areas on the surface of the eyeor hydrophobic interactions with the hydrophobic regions of the eye thatplay a role in dry eye syndrome. Similarly, these polymers couldinteract with natural mucins and be effective in adhering to mucinsand/or keeping soluble mucins in the tear longer, and/or increasing theeffectiveness and natural protections provided by mucins. Theformulations and use of the graft and block copolymers described hereinmay also have beneficial effects based on the steric stabilization ofbiological surfaces. The protection provided may help break the viciouscycle of inflammation, cellular injury, and discomfort.

Formulations comprising a bi-functional co-polymer and an anestheticagent such as proparacaine can be used in accordance with the presentinvention to prolong the anesthetic effect and/or reduce the acutecorneal surface changes seen commonly in the clinic and operating roomassociated with decreased sensation of the cornea and a decreased blinkrate. Such formulations can be applied prior to examination and/orpre-operatively, and will help maintain a more normal corneal epithelialsurface. Furthermore, use of the formulation will help reduce bacterialadherence at sites of any procedures performed on the eye and will helpreduce post-procedural infection.

Formulations described herein also provide a safe, nonirritatingexcipient for ophthamological formulations with one or more additionalactive pharmaceutical ingredients. For example, one aspect of theinvention is the use of the block and graft copolymers described hereincombined with active agents for treating ophthalmic disease. Theirinclusion may reduce the irritation or tachyphylaxis (associated withsome eye drops), or simply provide an additional lubricating andwettable ocular surface enhancing the comfort and acceptance of eyedrops. Exemplary additional active pharmaceutical ingredients forophthamological uses include, but are not limited to, lubricants anddemulcents, as described supra, antibiotics (fluoroquinolones,vancomycin, cephalosporin, gentamycin, erythromycin, azithromycin, sulfadrugs, bacitracin, gatifloxacin, levofloxin, moxifloxacin, ofoxacin),acetazolamide, antazoline, aspirin, atropine, azelastine, bacitracin,betaxolol, bimatoprost, botanical drugs including zeaxanthine lutein,lycopene brimonodine, brinzolamide, carbachol, carteolol, ciprofloxacin,ofloxacin, cromalyn, cyclosporine, cyclosporine pro-drugs andcyclosporine derivatives, other immunomodulators, dapiprazole,dexamethasone, diclofenac, dipivifren, dorzolamide, epinephrine,erythromycin, fluoromethalone, flurbiprofen, gentamycin, glaucomamedications (prostaglandins, carbonic anhydrase inhibitors, epinephrineor alpha-agonists, beta-blockers), gramicidin, homatropine,hydrocortisone, hyoscine, keterolac, ibuprofen, ketotifen, latanaprost,levobunolol, levocabastine, levofloxin, lotepprednol, medrysone,methazolamide, metipranolol, naphazoline, natamycin, nedocromil,neomycin, neuroprotective agents, nonsteroidal anti-inflammatory agents,nepafanec, norfloxacin, ofloxacin olopatadine, oxymetazoline,pemirolast, pheniramine, phenylephrine, pilocarpine, povidone,prednisolone, proparacaine, scopolamine, tetracaine, steroids,sulfacetamide, tetrahydrozoline, hypertonic tears, timolal, tobramycin,travaprost, trifluridine, trimethiprim, tropicamide, unoprostone andzinc. Prodrugs and related compounds, as well as any new activepharmaceutical ingredients can be used with the block and graftcopolymers here described.

Formulations comprising a graft and/or block co-polymer can also be usedin accordance with the present invention with contact lens solutions,manufacturing, rewetting drops, or in pre-insertion contact lenstreatments. The graft and/or block co-polymers can be used with currentcontact lens care solution and rewetting drop ingredients, including,but not limited to, water, preservatives, NaCl and other salts andmineral ingredients, buffers, and other polymers and osmotic agents. Theuse of these graft and block co-polymers in contact lens relatedconsumer products to impart clinical benefits is claimed. Also claimedis the use of these graft and block copolymers in the manufacture,coating, finishing, and storage of contact lenses.

Formulations comprising a bi-functional co-polymer can also be used inaccordance with the present invention with a second eye drop, such as alipid or oil based eye drop, to enhance the efficacy of the second eyedrop. In one embodiment of this use, the formulation described herein isadministered first followed by administration of the second eye drop.

Additionally, these formulations with the ability to prevent or inhibitcell adhesion are useful in preventing or inhibiting the spread ofbacterial or viral infections such as bacterial (including Chlamydial,gonorrheal, Staphylococcus epidermidis, Streptococcus pyogenes,Streptococcus pneumonia, Neisseria meningitidis and Moraxella lacunata,Haemophilus and family Enterobacteriaceae, Clostridium species,gram-negative anaerobic bacilli, and Peptostreptococcus species, Ngonorrhoeae, C trachomatis, Staphylococcal species and S pneumonia) orviral keratitis or conjunctivitis (including herpetic keratitis andadenovirus conjunctivitis) and in preventing and/or inhibitinginflammation caused by adherence of inflammatory cells and proteins tothe eye surface. The invention may be useful in treating or preventingamebic, protozoal, mycobacterial and other types of keratitis andconjunctivitis. There may be a benefit in treating sterile ulcers,ectasia, and corneal or scleral melts. The anti-inflammatory effect mayalso be due to inciting agents being kept off the corneal andconjunctival surface decreasing the inflammatory reaction. Suchformulations will be particularly useful in neonatal, daycare, pediatricor family settings wherein outbreak of one individual can lead toinfection of many others.

For the eye, the application of a formulation in accordance with thepresent invention is expected to reduce the exposure of the surface ofthe eye to bacterial antigens, such as those from Staphylococcus aureus,and thus may provide an important part of the treatment for staphmarginal disease. There also may be benefits to patients with allergicconjunctivitis including giant papillary conjunctivitis (limiting directtissue exposure to antigens), and for patients who have had previousfiltering surgery with thin ischemic blebs, as therapy with aformulation in accordance with the present invention is expected toreduce the risk of bleb related endophthalmitis. Additionally, after acorneal abrasion application of a formulation in accordance with thepresent invention can decrease the risk of corneal ulcer development,especially in the setting of injuries sustained from vegetable matterwhich predispose a subject to fungal infections. Other conditions thatcould be treated with formulations described with this invention includeconjunctivitis and keratitis due to processes such as Moorens ulcer,Terriens marginal degeneration, ligneous conjunctivitis, toxicexposures, autoimmune conjunctivitis, and phlyectenular conjunctivitis.Use in patients with keratoprostheses is claimed. The eye drops can beused in mammals, humans, or even dogs, cats, and horses.

From experiments described herein, it is expected that formulations ofthe present invention will also be useful changing wettability and/or,steric stabilization, tribological properties, lubricating and/orpreventing adherence of unwanted proteins and cells to other biologicalsurfaces including, but not limited to, skin, mucous membranes and hair.These formulations can thus also be applied in accordance with thepresent invention to epithelial tissue of the skin, urinary, orgastrointestinal tract, mucous membrane, exposed wound surfaces,including respiratory tract mucosa, oral and nasal mucosa, vaginalmucosa, and conjunctival surfaces, and surgical and traumatic wounds andulcerations. These formulations can serve to protect skin and otherorgans from foreign protein, viral, and bacterial adherence. Benefitscan include reduction in rate of infection. Without being limited to anyparticular mechanism, it is believed that a formulation comprising agraft polymer as described above, such as PLL-PEG, adheres to thebiological surface via the PLL moiety while the PEG moiety preventsand/or reduces the adherence of potentially harmful particles. Decreasedadherence can be beneficial by decreasing pathogen or toxin exposure,thereby decreasing morbidity and mortality associated with these agents.For example, lower bacterial loads can decrease the severity ofsubsequent bacterial infection and/or exotoxin exposure, and allow thehost defenses and antibiotics better opportunity to work. Decreasingviral exposure can reduce transmission rates and possibly the severityof viral infection. These formulations are useful as well againstfungals and exposure to reactive proteins.

Accordingly, the formulations described herein have a wide variety ofuses in accordance with the present invention.

For example, these formulations can be used in accordance with presentinvention in military applications to reduce soldiers' and citizens'morbidity associated with biological or biochemical warfare attacks.

These formulations can also be used in accordance with the presentinvention to reduce rates of methicillin-resistant Staphylococcus aureus(“MRSA”) adherence in hospitalized patients, children, and the generalpopulation. MRSA is a growing concern in the health care setting.Reducing the adherence to the epithelial surface thereby reducing MRSAspread and severity and frequency of MRSA infections is expected toprove very beneficial. Such use is expandable to school situations,prehospital admission, nursing homes and chronic care facilities aswell. It is not expected that MSRA, or other pathogens, would developresistance to formulations of the invention.

These formulations can also be used in accordance with the presentinvention to reduce transmission of the common cold and influenza virusvia nasal and inhalational applications in settings including, but in noway limited to, airplanes, preschools and schools, homes of affectedviral individuals, as well as nursing homes and chronic care facilities.The block or graft co-polymers can be included in nasal sprays andproducts formulated for inhalational delivery, as is known to thoseskilled in the art. For example, a nasal spray would include the blockor graft co-polymers and benzalkonium chloride, dextrose anhydrose,edentate disodium, microcrystalline cellulose and carboxymethylcellulosesodium, polysorbate 80, and purified water. An aerosol inhaler wouldinclude the block or graft co-polymers as well as ethanol and propellantsuch as propellant HFA-134a (1,1,1,2-tetrafluoroethane).

Important utilities of the formulations exist for children and adultswith infirmities, such as immunodeficiencies, chronic disease and otherchronic conditions such as cystic fibrosis, which leave the host moresusceptible to routine illness.

Application of a formulation in accordance with the present invention toa wound just after injury or surgery, including, but not limited tocorneal abrasions, corneal surgery such as LASIK, PRK, or otherrefractive or vision correction procedures, or intraocular surgery wherean incision is made in the eye through sclera or conjunctiva/sclera,such as for cataract or glaucoma surgery. The formulation can be usedagain after cleansing, can reduce bacterial infection, and infectiouswound complications. Such formulations may be particularly useful in thefield in military medical operations, after an initial irrigation.

For these uses in humans, the formulation may be in the form of alotion, gel, liquid, spray, rinse, dissolvable wafer, or glycerin bar towhich water is added to solubilize the graft co-polymer or blockco-polymer to make it more amenable to application. Formulations can beprovided as individual or single use products or in volumes forindustrial use and/or multiple use dispensers. In addition to thebi-functional co-polymer, such formulations may comprise any and alltypical binders, excipients, and components found in cosmetic sprays,lotions, soaps, shampoos, cleansers, and oral, nasal, vaginal, and eyecare products.

Formulations of the current invention can be delivered by eye dropbottles with a nozzle on the end (volume generally between 0.2 cc and 50mL). The container may be single use disposable containing a volume ofapproximately 0.3 mL or a multi use vial containing a volume between 0.3mL and 50 Ml. The material would be high density polyethylene orsimilar, and the container could be up to 500 mL for multipurposecontact lens solutions, and would have a nozzle. The solution could bestored in contact lens containers, as well.

By “eye care solution container” it is meant a container for eye caresolutions that is made of high density polyethylene or similarthermoplastic or petroleum based plastics for health care use. The eyedrop bottle is made from a mold or by blow, fill and seal methodology.The eye drop solution container has a nozzle for releasing smallerquantities of the solution than the total volume of the container. Eyecare solution container may be single use or multi-use. The volumes foran eye care solution container range from 0.1 cc to 500 cc.

Formulations comprising a bi-functional co-polymer with a positivelycharged or hydrophobic moiety and a hydrophilic moiety can also be usedin accordance with the present invention in the treatment of dry mouthin subjects with, for example, but not limited to, Sjogrens syndrome, ordry mouth during or post chemotherapy. Such formulations may furthercomprise traditional lubricants for such dry mouth conditions, such as,but not limited to, xylitol, and glycerin MouthKote.

The ability of these formulations to lubricate mucous membranes withoutirritation makes these formulations useful as sexual lubricants as well.Reduced transmission of HIV and other sexually transmitted diseases whena formulation is applied prophylactically on skin and mucosal surfacesin accordance with the present invention prior to exposure may be anadditional, if not a primary benefit in this application.

Formulations can also be used in accordance with the present inventionto protect gums in, for example, periodontal disease by reducinginflammatory cell and bacterial cell adherence to the gums. In thisembodiment, these formulations may be incorporated into, for example,but not limited to, a toothpaste or mouthwash and may be combined withflavorant and/or fluoride. Formulations which reduce bacterial adherencein the mouth, and in particular the posterior tongue and throat, arealso expected to be useful in treating bad breath or halitosis.

Formulations can also be used in accordance with the present inventionas a preoperative shampoo to reduce bacterial adherence to hair and/orskin before surgery. Since these formulations have been found to be safefor administration to the eye, they provide a useful preoperativeshampoo to remove any bacteria adhering to the eyelashes prior to eyesurgery.

The ability to reduce or prevent bacterial adherence to biologicalsurfaces such as skin and hair is expected to make these formulationsuseful in the treatment of chronic blepharitis as well, with or withoutconcomitant steroids or antibiotics. Formulations described herein maybe used on the eyelids and eye lashes, for treatment of medicalconditions and for prophylaxis. For these embodiments, the formulationmay be combined with an antiseptic or an antibiotic.

Further, the ability to reduce bacterial adherence to skin makes theseformulations useful in deodorants to reduce body odor caused by bacteriathat break down sweat.

Formulations can also be used in accordance with the present inventionon animals, including household pets, to decrease bacterial adherence totheir skin, oral cavity, or coats. In this embodiment, the formulationcan be in the form of a spray, shampoo, lotion, or beverage.

Other uses for these formulations will become evident to those skilledin the art upon reading this disclosure and are such uses areencompassed by the present invention.

The present invention also provides coatings comprising a graftco-polymer with a positively charged or hydrophobic moiety and ahydrophilic moiety or a block co-polymer with a positively charged orhydrophobic moiety and a hydrophilic moiety for extraocular devices suchas contact lenses, glaucoma stents and valves, scleral buckling hardwareand external drug delivery devices. External drug delivery devices caninclude biodegradable devices as well as permanent products such asthose made of silicon and other non bioerodible polymers. There is apotential use for intraocular devices including, but not limited to,intraocular lenses, intraocular contact lenses, glaucoma stents, tubesand valves, intracorneal implants that can be used for the correction ofpresbyopia, intraocular pressure sensors (including devices forcontinuous or microchip measurements with or without telemetry forintraocular pressure and intracranial pressure monitoring), andmicrochips used for retinal vision enhancement. Such coatings improvewearability and biocompatibility of these extraocular devices (such ascontact lenses) and decrease proteinaceous, bacterial, fungal, andparticulate adhesions to these extra-ocular devices. These coatings alsoimprove optical performance of the lenses. Coating intraocular andextraocular devices in accordance with the present invention alsoreduces epithelial adhesion and fibrosis thereto.

For example, contact lenses are often difficult to tolerate due to thediscomfort caused by the adhesion of proteins such as lysozyme anddenuded protein products and other particles to the negative charges ofthe lens surface. Other particles adhering to the lens surface includebacteria and bacterial proteins, endotoxins, antibodies, calcium, andlipids. Other uncharged particles, and even negatively charged particlescan build up on the lens surface once this cascade of protein adhesion,cellular and/or particulate adhesions to the lens is initiated. Wearingof contact lenses with these materials adhered to the contact lens canlead to discomfort, allergic reactions, ocular irritation, foreign bodysensation, epithelial breakdown and dysfunction, and bacterialkeratitis.

By binding the surface of a contact lens with positively charged and/orhydrophobic domains of block or graft co-polymers included in topicalformulations of the present invention, the aggregation of proteindeposits during wear time on the lens is and will be reduced and contactlenses are and will be better tolerated. The lens surface is also morelubricious, and the steric stabilization as well as high surface waterretaining capacity and charge neutrality can impart numerous benefits inthis clinical setting. With protection provided by the polymers hereindescribed, the contact lenses may also be more wettable and feel morecomfortable with the concomitant use of formulations described herein aswell as other ophthalmic products. Formulations can be provided inaccordance with the present invention as eye drops to improve contactlens wearability, as a finishing rinse after lens manufacture, instorage solutions, in lens cleaning solutions, in rewetting drops, as adrop to be used prior to placing the lens in the eye, and/or as anactive preservative in contact lens preparation and storage. Theseformulations thus may be used in accordance with the present inventionin the form of a one time application or repeated application. Contactlenses and intra- and extra-ocular devices can be made morebiocompatible with the coatings of the graft and block co-polymers asdescribed herein.

There are well known similarities, and there exists an establishedclinical relationship between dry eye syndrome and contact lensintolerance. In studies described in examples 4 and 5 in this invention,it has been shown clinically that the formulations and uses of graft andblock copolymers described herein conferred meaningful and measurablebenefit in dry eye subjects. These results are highly suggestive of asignificant opportunity for clinical benefit in the contact lens use andcontact lens intolerance clinical setting. By way of interpolation, itfollows that there will be similar benefits conferred in the use ofthese graft and block copolymers on other extraocular- and intra-cornealdevices.It can be foreseen that some intra-corneal or extra-ocular devices willhave prolonged biological use (for example extended wear contact lens,and surgically implanted devices). Under conditions of high stress (suchas ionic, salts or other electrolyte media, enzymatic, pH, ortemperature) and likely prolonged time in aqueous electrolyte media, theelectrostatic or hydrophobic interactions between the graft and blockcopolymers and the intra- or extra ocular devices may be reduced to somedegree. Additionally, the tear film, sub-epithelial, or intraocularfluid will likely not (unless a specific intervention has beenundertaken for that purpose) be a reservoir for replacement graftcopolymers as they naturally degrade over time. The benefit conferred bythe polymers to the device may, therefore, wear off over time. Thus,there may be advantages to covalently immobilizing the block or graftcopolymers herein described to the intra or extra ocular device duringmanufacturing or finishing. It follows, therefore, that covalentlybonding, embedding during manufacture, or covalently immobilizing thesegraft and block co polymers to the devices may be beneficial. It hasbeen shown that covalent immobilization does not necessarily result inless efficient packing of a graft co-polymer (Blattler, et al. Langmuir2006, 22, 5760-5769). An aldehyde plasma polymer interlayer withreductive amination can be used in such a setting. Other methods forcovalently immobilizing the block or graft copolymers can alsoaccomplish the same result of developing a more stable and resistantcoating on the surface of the device.

Use of block or graft co-polymer containing formulations to coatintracorneal devices also reduces particulate, cellular, epithelial,and/or fibrocyte adhesion. Intracorneal inlays, may benefit from the useof these graft and block copolymers. Reductions in the risk of infectionand clotting or blockage by protein or fibrin debris, cellularproliferation, inflammation suppression, and steric repulsion are allways in which these polymers may confer advantages to extra- orintraocular devices, including intracranial implants.

The descriptions provided above are not to

be construed as limitations of the present invention and its variousembodiments described herein, but rather, are described to fully allowone skilled in the art to make and use the invention along withmodifications, adaptations, and alterations, all being within the spiritand broad scope of the present invention as fully defined herein.Similarly, the following nonlimiting examples are provided to furtherillustrate the present invention. Percentages are weight percentages.

EXAMPLES Example 1

The following topical formulation was made and tested:

-   1% F127-   1% PLL-g-PEG-   1% glycerin-   sterile water for injection-   sodium phosphate buffer-   mannitol to adjust osmolality;-   pH between 6.5 and 7.5 and osmolality 274-350 mOsm/kg;-   Sterile filtration

This formulation was tested in the Draize test and no ocular irritationwas observed.

This formulation was also tested in two volunteer humans and noirritation was reported. Tear film break up time in one volunteer wasextended compared to the control eye for over two hours afterinstillation of the eye drops. On close inspection under microscopy,after instillation, the tear film of the eye treated with the PLL-g-PEGformulation displayed a more stable tear film. This stability waspresent at greater than three hours after eye drop instillation.Furthermore on analysis using a Tearscope to evaluate the tear film,compared to an eye that received a commercially available product, theeye treated with the PLL-g-PEG polymer showed an increase in tear filmbreak up time by 30 seconds compared to control, on one occasion, at onehour after instillation of the eye drops.

This formulation was also applied to a contact lens and tested. One dropwas placed on each side of a contact lens. The lens was placed in ahuman eye and the patient wore the contact lens comfortably for 8 hours.The treated lens was described as being more comfortable in the eye thanan untreated control lens. This formulation was also mixed 1:1 with acommercially available preserved multi-purpose contact lens solution andthe lens was stored in the solution overnight. The lens was worn thenext day comfortably. Again, the treated lens was more comfortable thanan untreated control lens. One embodiment of the copolymers hereindescribed has thus been used as an eye drop, used in combination withother polymers, used with contact lenses, and formulated with apreservative. The subject felt decreased awareness of the contact lensand less resistance and discomfort when blinking.

Example 2

An in vitro assessment was made of the ability of 1%, 0.5%. and 0.1%PLL-g-PEG formulations and 1% formulation of F127 and F87 to decreasecontact angle of water on a polystyrene surface. The contact angle of adrop of these had a much lower contact angle on a freshly cleanedpolystyrene surface compared to a drop of deionized water. Thisexperiment demonstrates the ability of these formulations to changewettability and adherence to negatively charged surfaces such as the eyeand epithelium.

Example 3

An ex vivo assessment in porcine eyes was made of the ability of aPLL-g-PEG formulation of the present invention to decrease evaporation.Porcine eyes were obtained fresh and refrigerated. They were stored inan oil bath, which caused changes to the epithelial surface and alsopartially denuded the epithelial surface allowing for faster evaporationof an aqueous solution. The evaporation rates of small areas weremeasured and Systane was applied with no change in evaporation rateafter a single rinse. A single rinse with the PLL-g-PEG formulationshowed a reduction in evaporation time by a mean of two seconds. Fiveeyes were tested. The formulation was: 1% F127, 1% PLL-g-PEG, 1%glycerin, sterile water for injection, sodium phosphate buffer, mannitolto adjust osmolality.

Example 4

Two tests were performed in human eyes with a PLL-g-PEG formulation ofthe present invention.

The formulation was: 1% F127, 1% PLL-g-PEG, 1% glycerin, sterile waterfor injection, sodium phosphate buffer, mannitol to adjust osmolality.

The first involved the time a human eye could be held open before theneed to blink based on discomfort. The longer an eye could be held openwithout pain implies a better tear film covering over time. In thisexperiment, one eye received a single eye drop of Optive™ while theother eye received a single eye drop of the PLL-g-PEG formulation eyedrop. The control eye could be held open 30 seconds after instillationwithout discomfort repeatedly at most time points (every 10 to 15minutes) from t=5 minutes to t=120 minutes. The eye treated with thePLL-g-PEG formulation could be held open without discomfort for timesranging from 40 to 60 seconds at most time points.

The tear film was also assessed by Fluorescein tear film break up time.At t=200 minutes after instillation, there was a longer time to TFBUT byat least 5 seconds in the PLL-g-PEG treated eye. Additionally, the breakup occurred with a much smaller area of change on the surface of theeye. These results were confirmed subjectively with the wavescan devicelooking at tear film reflectance of light and aberrations.

Example 5

The formulation was: 1% F127, 1% PLL-g-PEG, 1% glycerin, sterile waterfor injection, sodium phosphate buffer, mannitol to adjust osmolality.

A randomized, masked, active comparator controlled study was carried outin eighteen subjects. The study endpoints included subjective responseto the eye drops and the tear film break up time compared topreinstillation values at t=15, 30, 60, and 120 minutes. Noninvasivetear film break up time (NIBUT) was performed using the Tearscope™ in 16subjects; fluorescein break up time (FBUT) at 120 minutes was alsomeasured in these sixteen subjects. Two subjects were tested withwavescan techniques to look at tear film break up time with that newtechnology. A questionnaire was administered to subjectively assess theacceptability of the eye drops in all subjects. The principalinvestigator was not affiliated with the product and is a full professorat an academic eye institute. The principal investigator was responsiblefor the trial. IRB approval was obtained, and the trial was registeredat clinicaltrials.gov, as per FDA requirements. The comparator was aleading over-the-counter product with the following formulation: ActiveIngredients: Polyethylene glycol 400 0.4% and propylene glycol 0.3%.Inactive Ingredients: boric acid, calcium chloride, hydroxypropyl guar,magnesium chloride, potassium chloride, purified water, sodium chloride,and zinc chloride. In the Tearscope portion with sixteen subjects, threegroups were studied: the first group of five subjects had none,occasional, or mild dry eye symptoms. The second group of five subjectshad mild to moderate dry eye symptoms. The third group of six hadmoderate to severe dry eye symptoms. RESULTS: Questionnaire: there wasno difference overall between the acceptance of the eye drops. See Table1.

TABLE 1 Preferred at 5 minutes after instillation Sample formulation 4Active comparator 5 No difference 7Vision changes: No change: 100% Eyeon; No change: 87.5% (2 said visionwas blurrier after active comparator was received. Comments at two hoursif any: Equivalent for active comparator and study formulation.Regarding slit lamp exam, there were no adverse events in any eyes, andthere were no changes in the anterior segment exam in any eyes. NIBUTresults for the entire cohort: 15 minute time point showed NIBUT timeincrease from baseline at plus 14.67 sec (p<0.05) for sampleformulation, while active comparator was 7.4 seconds longer thanbaseline (p=0.3). Thus, the new formulation essentially doubled the tearfilm break up time benefit of the active comparator at 15 minutes. FBUTwas significantly longer than active comparator at 120 minutes; superiorby 4.92 sec (p<0.12). See FIGS. 3 and 4.The two subjects who had wavescan evaluation showed results thatsupported the conclusions in the larger study, for example, the meanTFBUT was longer for the sample formulation than active comparator atall time points.Furthermore, a sub group analysis was performed. In the main group of 16subjects there was one subject in each group that had unexpectedly longpre-instillation TFBUT. These long initial values distorted the relativecontributions of the artificial tears to NIBUT for the other subjects.When these three outliers were removed, there was superiority inextending NIBUT for the sample formulation at every time point. The datawas evaluated by seconds over baseline, and by percentage change frombaseline. See FIGS. 5 and 6.Thus, this randomized, controlled, masked trial supports use of theinvention for treatment, including prevention, of dry eye, contact lensintolerance, ocular irritation, and other disorders of the skin andmucous membranes.

Example 6

There is a need for compositions for treating injuries that can becustom loaded with therapeutic agents based on the suspected bacterialspecies in a wound on an external body surface (e.g., the skin).Non-restrictive loading could potentially reduce bacterial colonizationby orders of magnitude and reduce infection rates in compromisedpatients with contaminated wounds. The following results indicate thatcompositions containing a copolymer having a positively charged, orhydrophobic, or covalent bonding moiety and a hydrophilic moiety areuseful as a carrier for antibiotics. Such compositions may be usedalone, as an adjunctive therapy to wound treatment, or to protectagainst pathogen or toxin attack (e.g., from bacterial infection).

A composition containing a copolymer having a positively charged, orhydrophobic, or covalent bonding moiety and a hydrophilic moiety isformed into a wound management device or formulated in a pharmaceuticalcomposition. The copolymer composition is tested on a mouse model ofwound healing. Mice treated with the copolymer composition have fasterand/or more complete wound healing compared to untreated mice. Micetreated with the copolymer composition also have fewer or less bacterialinfection compared to untreated mice.

The copolymer composition is loaded with the antibiotic amikacin.Amikacin release from the copolymer composition is measured. The openwounds of the mice may be passively or actively infected with bacteria.Bacterial growth inhibition of P. aeruginosa is observed for woundstreated with the copolymer composition. Treatment of wounds with acopolymer composition with antibiotics results in inhibition ofbacterial growth and progress in wound healing compared to mice treatedwith the copolymer composition alone or with no treatment.

The ability to customize the antibiotic choice is advantageous becauseit allows clinicians to tailor treatment regimens based on known orsuspected bacterial species present. Copolymer compositions of theinvention allow for high release concentrations of antibiotics. Theseexperiments indicate that incorporation of antibiotics into a copolymercomposition provides a local drug delivery system that can be used aloneor in conjunction with wound treatment therapies.

Example 7

There is a need for nanoparticle compositions for treating nasal andinhalational conditions that can be custom loaded with active ortherapeutic agents based on the treatment required. Nanoparticles coatedwith a copolymer having a positively charged, or hydrophobic, orcovalent bonding moiety and a hydrophilic moiety can be used to deliveractive or therapeutic agents for inhalational applications. Forinhalation, the nanoparticle size should be 10 microns or smaller indiameter, about 1 micron or less in diameter. The copolymer coatednanoparticles are formulated with a pharmaceutically acceptable carrier.The copolymer coated nanoparticles can be used in a pharmaceuticalcomposition that is administered to the oral, nasal, or respiratorysurfaces (e.g., in the throat) using an aerosol or nasal formulation(e.g., a spray). The copolymer coated nanoparticles contain active ortherapeutic agents, including drugs (e.g., an antibiotic), menthol, orother numbing compound for treatment of coughs or sore throats. Thecopolymer coated nanoparticles deliver the active and therapeutic agentswhen contacted to oral, nasal, or respiratory surfaces (e.g., mucousmembranes). Thus, the copolymer coated nanoparticles are used to deliveractive and therapeutic agents.

Example 8

There is a need for an eye drop that helps the eye retain heat. Aninvestigation was carried out into the effect of the multi-functionalgraft polymer with a positively charged backbone and hydrophilic sidechains on the body's surface temperature at the cornea. The finding thattopical formulations of this the multi-functional graft polymer increasethe ocular surface temperature forms the basis of the instantlydisclosed new method for treating human eye conditions. Furthermore, theresults of the assays described in this example were demonstrativedirectly on the effect on temperature, not indirectly by timingevaporation on an isolated surface. The in vivo dynamics are complex dueto multiple variables including ocular blood flow, variabilities in airflow on a biological surface, and natural tear secretion. Thus, it wasdecided to independently measure temperature using thermal imaging ofthe surface of the eye to determine if the tear could also affect thesurface temperature of the eye. The stabilization of the tear film usingtraditional methodology such as fluorescein tear break up time has beendemonstrated above. It had never been shown that the formulations testedpreviously could have any measurable effect on temperature. Toward thatend a proof-of-principal study was undertaken, as described below.

Purpose:

To demonstrate if the test drop(s) has any effect on thermal imaging ofthe cornea by an infrared camera with concurrent assessment of the tearfilm using placido disc imaging.

Methodology:

One eye (left or OS) was measured at baseline placido disc and thermalcamera. Then in that eye, a formulation of 1% PLL-g-PEG, 1% glycerin, 1%F127, sterile water for injection, sodium phosphate buffer, mannitol toadjust osmolality was instilled. That eye was imaged (OS) with theplacido disc and thermal camera.Four different time points were studied—baseline (before dropinstillation), 5 min, 15 min and 30 min after drop instillation. Theairflow and environment was controlled and was the same for allmeasurements. The eye was examined prior to eye drop instillation and atthe time points indicated.The placido images were studied subjectively to assess tear break uptime (TBUT). Thermal images were also analyzed using custom MATLAB(matrix laboratory) software to compare average ocular surfacetemperature in the central 9 mm corneal region.

Results:

TABLE 2 Thermal Imaging Placido (Avg temp ± SD)° C. Baseline TBU at 8secs 33.97 ± 0.23  5 min TBU at 20 secs  34.4 ± 0.04 15 min No TBU seenuntil 35 secs, nice 35.04 ± 0.11 placido pattern, relatively easier tohold eye open 30 min No TBU until 38 secs, nice placido 35.05 ± 0.08pattern, no distortions noticed, relatively easier to hold eye open

Major Points—

-   -   The tear break up time by non invasive placido imaging increased        “significantly” from baseline to 5 mins. (significant tear film        break up time can be considered greater than 25% increase in        this example).    -   The break up time from the 5 minute time point to the 15 minute        and 30 minute time points increased substantially.    -   The ocular surface temperature at baseline was 33.97±0.23° C.        OST increased from baseline to 5 mins, and a marked increase is        noticed from baseline to 15 mins and 30 mins, by more than 1° C.        For this example, significant temperature increase is 0.34        degrees Celsius (1% increase).

Conclusions:

The test drop has a positive effect in increasing the ocular surfacetemperature. The test drop also increased the stability of the tear filmas demonstrated by the increased tear break up time. The eye toleratedthe test drop well with no symptoms or hyperemia.

Example 9

Example in contact lens/multi-functional polymer surface temperatureeffects. Based on the results obtained in Example 8, above, it can beshown that the use of a contact lens manufactured with or soaked inPLL-g-PEG or alternative (multifunctional graft copolymers) stabilizesor increase ocular surface temperature. For the experiment, the polymeris delivered via an eye drop, as a rewetting drop following theplacement of a contact lens. In this example, the contact lens is placedand after the eye equilibrates for greater than 5 minutes the ocularsurface temperature is measured on bulbar conjunctiva and on the contactlens external surface. The graft co-polymer is instilled. The surfacetemperature at either the bulbar conjunctive or the exposed surface ofthe contact lens is higher following instillation of the graftco-polymer with the lens in the eye as compared to the pre-instillationlens in the eye temperature measurements.

Example 10

Formulations without pluronics or active demulcents. To further clarifythe value of the graft-copolymer effect alone, and as supportiveevidence that the benefits conferred from topical formulations with themulti-functional polymer do not require other additives, the stabilityof the tear film was assessed in a subject using a composition thatincluded PLL-g-PEG in solution with neither a demulcent nor a PLURONIC.The composition was: 1% PLL-g-PEG, sterile water for injection, sodiumphosphate buffer, sodium chloride, and mannitol to adjust osmolality. Anexpert examiner objectively evaluated the stability of the tear filmusing fluorescein tear film break up time (FTBUT). The left eye of asubject, directly before instillation of the test eye drop, showed anine second FTBUT. Several minutes were allowed to pass so thefluorescein washed out of the eye. At that point, a single drop (50microliters) of the topical of ophthalmic solution containing 1%PLL-g-PEG but without the demulcent or PLURONIC was instilled in theleft eye. Exactly 15 minutes later the FTBUT was measured, and it was 16seconds. Clinical success of prolongation of tear stability wasdemonstrated.

TABLE 3 Clinical evaluation of FTBUT with topical ophthalmic formulationof 1% PLL-g-PEG without a demulcent or PLURONIC: Pre-instillationbaseline Post-instillation FTBUT at 15 FTBUT minutes Increase 9 seconds16 seconds 78%Thus, the multi-functional graft copolymer is responsible for thebenefits obtained from contacting the surface of the eye with solutionsthat contain other additives, actives, or excipients.

Example 11

Utilizing the multi-functional graft co-polymer with other actives inophthalmology. PLL-g-PEG was added to a commercial ophthalmic solutionknown as Combigan®. The amount of liquid in a single multi-use eye dropbottle of Combigan® containing the active ingredients of brimonidinetartrate 0.2% and timolol 0.5% was poured out into a sterile vessel andweighed. Using a calculation to create a 2% weight/weight solution withPLL-g-PEG polymer, the proper amount of 99% pure PLL-g-PEG was dissolvedinto the solution. The new formulation was sterile filtered and returnedinto the internally sterile eye drop bottle. The new formulation createdwas thus a combination glaucoma product with the addition of the graftcopolymer PLL-g-PEG. Tear break up time of the PLL-g-PEG formulation wascompared with an unchanged brand new Combigan eye drop: Combigan® withPLL-gPEG 2% vs. Combigan® alone.

TABLE 4 OD OS Tear film break up time Fluorescein, no anestheticBaseline measure 1 9.3 13.6 measure 2 13 12.3 measure 3 12.5 14 average11.6 13.3 eye drop instillation OD received Combigan ® with 2%PLL-g-PEG; OS standard Combigan ® 25-30 minutes post instillation,masked observer measure 1 29.94 12.1 measure 2 26.44 9.58 measure 329.91 23.11 average 28.76333 14.93 prolongation of tear film break uptime OD 17.16333 sec OS 1.63 sec IOP at 2 hours post instillation OD 8mmHg OS 8 mmHg Notes per examining MD: “The tear film OD looked morestable, more uniform, and the rate of tear break-up, once begun, wasless complete and of smaller diameter.”This experiment thus demonstrated and confirmed several key results.First, this study re-affirmed that the effect on tear stability is fromthe addition of the graft co-polymer only. Second, the addition of thispolymer did not affect the activity of the drug product actives onreducing the intraocular pressure in the subject. Third, the simplicityof formulation with the polymer was demonstrated. Another aspect of thisexperiment was the use of the corneal topographer to assess longstandingeffect of the polymer. Eighty minutes after eye drop installation inboth eyes, the Placido % read parameter which was reported out by themachine's algorithm when the eyes were held open for 30 seconds was97.50% OD and 87.30% OS. When the eyes were held open for 60 secondsvalues were 97.80% read OD and 93.20% OS. Thus, the corneal topographercan be used to show the beneficial effect on the corneal surface qualityimparted by this PLL-g-PEG formulation, and be inference amulti-functional graft co-polymer with a positively charged backbone andhydrophilic side chains.

Example 12

Eyelid margin inflammation is treated with a multifunctional graftcopolymer. The eyelid margin temperature is 34 degrees Celsius beforetreatment. After application of an eye drop with a multifunctional graftcopolymer, the eyelid margin temperature increases to 35 degreesCelsius. After daily treatment for one week, the eyelid margininflammation is reduced.

Example 13

A patient had a non-healing leg ulcer from poor peripheral perfusion.The temperature on the surface of the ulcer is 34 degrees Celsius. Theulcer is treated topically with a formulation containing amultifunctional graft copolymer. Post application, the ulcerstemperature increases to 34.5 degrees Celsius. Over several weeks withdaily application, significant improvement has occurred. This change intemperature helps heal the ulcer faster as more blood flow and a lowerrisk of infection lead to better outcomes.

What is claimed is:
 1. A method to increase the surface temperature onthe external surface of a treated contact lens that is on the eye of asubject as compared to an untreated contact lens that is on the eye of asubject, comprising contacting the eye of said subject with said treatedcontact lens, wherein said treated contact lens is in association with apharmaceutical composition comprising a copolymer having a positivelycharged moiety and a hydrophilic moiety, wherein said contacting iseffected in an amount and for a duration so as to increase the surfacetemperature on the external surface of said treated contact lens that ison the eye of said subject as compared to an untreated contact lens thatis on the eye of said subject, thereby increasing the surfacetemperature on the external surface of a treated contact lens that is onthe eye of a subject as compared to an untreated contact lens that is onthe eye of a subject.
 2. The method of claim 1, wherein said contactingthe eye of said subject with said treated contact lens increases thetemperature of the ocular surface of the eye of said subject in contactwith said treated contact lens.
 3. The method of claim 1, wherein thecopolymer is a graft copolymer.
 4. The method of claim 1, wherein thecopolymer a block co-polymer.
 5. The method of claim 1, wherein thehydrophilic moiety is one of non-ionic or anionic.
 6. The method ofclaim 1, wherein the copolymer creates an electrostatic adhesion betweenthe copolymer and the biological surface.
 7. The method of claim 1,wherein the copolymer creates a covalent adhesion between the copolymerand the biological surface.
 8. The method of claim 1 wherein thecopolymer creates an electrostatic adhesion with the contact lens. 9.The method of claim 1 wherein the copolymer creates a covalent adhesionwith the contact lens.
 10. The method of claim 3, wherein the copolymeris a graft copolymer and comprises a cationic backbone and side chainsthat are water soluble and non-ionic.
 11. The method of claim 3, whereinthe copolymer is a graft copolymer and comprises a water solublenon-ionic backbone and cationic side chains.
 12. The method of claim 1,wherein the copolymer is a block copolymer and comprises at least onecationic block and at least one block which is water soluble andanionic.
 13. The method of claim 1, wherein the copolymer comprises0.001 to 40% of said composition.
 14. The method of claim 1, wherein thecopolymer comprises 0.001 to 25% of said composition.
 15. The method ofclaim 1, wherein said composition further comprises a second polymer.16. The method of claim 1, wherein said composition further comprises aPLURONIC block copolymer.
 17. The method of claim 1, wherein saidcomposition further comprises one or more of a surfactant, apreservative, and a pharmaceutical ingredient selected from the groupconsisting of: demulcents, emollients, sodium chloride, andvasoconstrictors.
 18. The method of claim 1, wherein said treatedcontact lens has been stored in said pharmaceutical compositioncomprising said copolymer having a positively charged moiety and ahydrophilic moiety.
 19. The method of claim 1, wherein said increase intemperature ranges from 0.1 degrees Celsius to 0.5 degrees Celsius. 20.The method of claim 2, wherein said increase in the temperature of saidocular surface of the eye of said subject in contact with said treatedcontact lens is at least 0.1 degrees Celsius.
 21. The method of claim20, wherein said increase in the temperature of said ocular surface ofthe eye of said subject in contact with said treated contact lens ismeasured at the conjunctiva.
 22. The method of claim 1, wherein saidincrease in temperature is measured using a method selected from thegroup consisting of ocular thermography, infrared thermal imaging,temperature distribution assessments, dynamic thermography, ocularsurface temperature assessment and ocular thermometry.
 23. The method ofclaim 1, wherein said increase in temperature is measured with athermistor or a thermal camera.
 24. The method of claim 22, wherein saidincrease in temperature is measured over time using a thermal camera,wherein said increase in temperature is measured in degrees Celsius. 25.A method to treat blepharitis in a patient by increasing the temperatureof the affected contact-lens wearing eye by treating said contact lenswith a copolymer having a positively charged moiety and a hydrophilicmoiety, wherein said contacting is effected in an amount and for aduration so as to increase the surface temperature on the externalsurface of said treated contact lens that is on the eye of said patientas compared to an untreated contact lens that is on the eye of saidpatient, thereby treating blepharitis in said patient.
 26. A method totreat a chalazion in a patient by increasing the temperature of theaffected contact-lens wearing eye by treating said contact lens with acopolymer having a positively charged moiety and a hydrophilic moiety,wherein said contacting is effected in an amount and for a duration soas to increase the surface temperature on the external surface of saidtreated contact lens that is on the eye of said patient as compared toan untreated contact lens that is on the eye of said patient, therebytreating chalazion in said patient.
 27. A method to treat dry eyesyndrome in a patient by increasing the temperature of the affectedcontact-lens wearing eye by treating said contact lens with a copolymerhaving a positively charged moiety and a hydrophilic moiety, whereinsaid contacting is effected in an amount and for a duration so as toincrease the surface temperature on the external surface of said treatedcontact lens that is on the eye of said patient as compared to anuntreated contact lens that is on the eye of said patient, therebytreating dry eye syndrome in said patient.